PRACTICAL PROJECT DIPLOMA OF PUBLIC SAFETY (FORENSIC INVESTIGATION) Submitted By: Brett Griffin

Transcription

1 Alternative Logo DIPLOMA OF PUBLIC SAFETY (FORENSIC INVESTIGATION) PRACTICAL PROJECT The feasibility of using ABAcard p30 Test Kits at a crime scene where it is alleged a sexual assault took place. Submitted By: Brett Griffin 2010

2 The feasibility of using ABAcard p30 Test Kits at a crime scene where it is alleged a sexual assault took place. A PROJECT SUBMITTED IN PARTIAL FULFILMENT OF THE REQUIREMENTS FOR THE DIPLOMA OF PUBLIC SAFETY (FORENSIC INVESTIGATION), AT THE CANBERRA INSTITUTE OF TECHNOLOGY. BY Brett Griffin 15 th November 2010 Certificate of Originality 1. I certify that the work in this report has not previously been submitted for a degree nor has it been submitted as part of requirements for a degree except as fully acknowledged within the text. 2. I also certify that the report has been written by me. Any help that I have received in my research work and the preparation of the report itself has been acknowledged. In addition, I certify that all information sources and literature used are indicated in the report. 3. I certify that, where required, I have received ethics clearance from the appropriate authorities in accordance with CIT policies on human and animal research. Signed: Date: 15 th November 2010 Prepared by Brett Griffin Page 2

3 Acknowledgments I wish to thank the following people for their assistance in this study. Without their assistance this study would not have been possible. Amy Davis, BAppChem, DipSciPrac, the Scientific Officer with Chemical Criminalistics, Forensic Science Services Branch, at the New South Wales Police Force Forensic Laboratory, Pemulwuy and her staff for the advice and assistance given in the preparation of the semen solutions used in this study. Senior Constable Graham Lovelace of Penrith Crime Scene section who assisted in sourcing substrates used during this research. Prepared by Brett Griffin Page 3

4 Table of Contents Abstract...05 Background...07 Materials and Methods...11 Products...17 Results Summary of Results.77 Discussion...79 Conclusion...85 Recommendations...86 References...87 Annexure...88 Prepared by Brett Griffin Page 4

5 Abstract The detection and identification of latent semen stains at crime scenes of a sexual nature is an integral part of the overall investigation. The New South Wales Police Force Forensic Services Group (NSWPF FSG) currently search for these latent semen stains using a combination of light sources and chemicals, however both of these techniques are presumptive tests only, and both of these techniques are subjected to false positives. The ABAcard p30 Test For The Forensic Identification of Semen at Crime Scene manufactured by Abacus Diagnostics, Inc. claims to be an confirmatory test for the presence of Prostate Specific Antigen (PSA), with no false positives associated with the currently used presumptive tests. This study directly compared the sensitivity of the ABAcard p30 Test Kit and the product which is currently in used by the New South Wales Police Force Forensic Services Group, the Phosphatesmo KM strips. The experiments were conducted over a four week period and used four substrates commonly found in sexual assault scenes, being carpet, timber floor boards, bed linen and ceramic tiles. Seven dilutions of semen were used throughout the study, being Whole semen, 1:10, 1:100, 1:1000, 1:10 000, 1: and 1: This study also evaluated other issues related to using products at crime scenes. These issues include Occupational Health & Safety (OH&S) requirements, how the products are stored, the transportation of the products to crime scene and the associated costs including purchase and training. The results of the study show that the ABAcard p30 Test Kit is an easy to use field confirmatory test. The ABAcard p30 Test Kit is at least as sensitive and in the majority of experiments substantially more sensitive then the Phosphatesmo KM strips. The ABAcard p30 Test Kit also has the added advantages of being a confirmatory test as well as having an inbuilt control system to prevent false positives. Prepared by Brett Griffin Page 5

6 The study has determined that the ABAcard p30 Test Kit has an overall higher sensitivity than the Phosphatesmo KM strips and with a built in control, the ABAcard p30 Test Kits would be a cost effective tool in the investigation of sexual related crimes. Prepared by Brett Griffin Page 6

7 Background Semen, as described in the Collins English Dictionary is: A viscous whitish secretion of the male reproductive organs, containing spermatozoa and consisting of secretions of the testes, seminal vesicles, prostate, and bulbourethral glands. Also called seminal fluid. Usually, each millilitre of semen contains millions of spermatozoa (sperm), but the majority of the volume consists of secretions of the glands in the male reproductive organs. Semen consists of the secretions of several glands but only 5 per cent comes from the testicles. (netdoctor.co.uk) Where semen is made Contributing gland Percentage of whole ejaculate Testicles and epididymes 5 per cent Seminal vesicles 46 to 80 per cent Prostate gland 13 to 33 per cent Bulbourethral and urethral glands 2 to 5 per cent Table 1: Where semen is made within the male body. Source netdoctor.co.uk Prostate-specific antigen (PSA) given the generic name p30, is a protein produced by normal prostate cells. This enzyme participates in the dissolution of the seminal fluid coagulum and plays an important role in fertility. The highest amounts of PSA are found in the seminal fluid. (Sensabaugh et.al.1978). The ABAcard p30 Test is an immunochromatographic 1 one-step tests for the detection of p30. If p30 is present in a sample, the p30 reacts with a mobile monoclonal antihuman 2 PSA antibody in the strip forming a mobile antigen-antibody complex. This antigen-antibody complex then migrates through the absorbent device toward the test area. A monoclonal antihuman PSA antibody is attached to the membrane in the test area. This immobilized antibody captures the above complex resulting in an antibody-antigen-antibody complex. When the p30 concentration is greater than 2-4 ng/ml, the dye particles will form a pink coloured band in the test area 1 an analytical method which uses antibodies as reagents. 2 a group of cells produced from a single ancestral cell by repeated cellular replication Prepared by Brett Griffin Page 7

8 indicating a positive result. The tests have an internal control to ensure that the test is properly working and that proper procedures have been followed. (Laux et. al., (2004). Being that the p30 antibody must be present in the same for the test area to produce pink coloured bands, the test can be used as a confirmatory test for the presence of p30. Figure 1 - A Positive (left) and Negative (right) ABAcard p30 test As a Forensic Investigator with the New South Wales Police Forensic Services Group (NSWP FSG), I often attend scenes of alleged sexual assault. At these scenes, it is essential to be able to accurately determine the presence or absence of semen. This is necessary to ensure that any semen that may be present is collected to be sent for DNA analysis. Currently, the method of semen detected used by the NSWP FSG is a presumptive based test using Phosphatesmo KM strips, where all stains that give a positive result are collected and sent to the Department of Analytical Laboratories (DAL) for confirmatory testing (New South Wales Police Force Forensic Services Group, 2009, Crime Scene Procedures Manual Specialist, section 21, Chemical Enhancement of Biological Evidence). Prepared by Brett Griffin Page 8

9 The NSW Police Force, Chemical Targeting of Trace Evidence course booklet defines a presumptive test as a test that is highly sensitive for a substance, but not specific for that substance (2009, p. 7). To put simply, a presumptive test for semen carried out by using Phosphatesmo KM strips is able to detect small quantities of semen, however it will also react with numerous other substances. This test method has its limitations as Phosphatesmo KM strips have numerous known false positive reactions, including but not limited to other bodily fluids such as vaginal secretions and saliva (Scott, 1998). The collection of exhibits following false positive reactions leads to unnecessary expenses to the NSWPS FSG, in that items that may contain semen evidence are being analysis at great expense and at the time of collection, even though you explain to investigators that it was a presumptive test, they still believe that they have located semen evidence at scenes. The product that I researched is the ABAcard p30 Test Kit manufactured by Abacus Diagnostics, Inc and sourced in Australia by XTEK Forensics. This research project examined the feasibility of the NSWP FSG using the ABAcard p30 Test Kits by determining its detection accuracy by using semen samples and by evaluating the ease of use in the field. This study concentrated upon semen of known dilutions that had been deposited upon various substrates commonly found within crime scenes. The sensitivity of both the currently used Phosphatesmo KM strips and the ABAcard p30 Test Kits was also directly compared. This study also looked at the practical application methods of both products and their results. Further, the study also examined other factors that impact on the two products including: Personal Protective Equipment required The shelf life of both products in both mixed and raw forms Storage and transport requirements The ability to record the results using digital photography Costs relating to initial purchase Availability of this product to the NSW Police Force Prepared by Brett Griffin Page 9

10 Training related issues This study took place over a period of four weeks and was conducted within the Penrith Crime Scene section examination room. Prepared by Brett Griffin Page 10

11 Materials and Methods Occupational Health and Safety Whilst conducting this research there were Occupational Health and Safety issues that were encountered. During all experiments the following were adhered to: The Occupational Health and Safety Act (2000) The NSW Police Forensic Services Standard Operating Procedures The Forensic Services Group Occupational Health and Safety Manual (2009) Whilst using the ABAcard p30 Test Kit I referred to the following: The manufacturer s instruction sheet Precautions section. The Forensic Services Group Occupational Health and Safety Manual (2009) Whilst using the Phosphatesmo KM TM strips I referred to the following: Relevant Material Safety Data Sheets (MSDS) The NSW Police Forensic Services, Chemical Targeting training module (2009) The Forensic Services Group Occupational Health and Safety Manual (2009) Prepared by Brett Griffin Page 11

12 Semen dilutions Fresh semen was used for all the experiments within this study. The following dilutions were made up with the assistance of Forensic Chemist, Amy Davies, BAppChem, DipSciPrac, Whole (neat) semen 1:10 1:100 1:1000 1:10,000 1:100,000 1:1,000,000 Figure 2: Rack of various semen dilutions used in this research Prepared by Brett Griffin Page 12

13 Surfaces and Substrates Numerous surfaces can be found within any crime scenes. This study was aimed at including surfaces that, from my own experience, suspected semen stains have been located upon. These surfaces are common to most homes. I used the following porous and non-porous substrates: Carpet (porous) Polished timber floor board (non-porous) Bed sheet (porous) Ceramic tile (non-porous) Seven circles were drawn on each substrate and labelled: Whole 1 : 10 1 : : : : : Each circle was then divided into three segments, to enable a sample to be collected from each circle three times at the following intervals: 3 days from application 2 weeks from application 4 weeks from application Prepared by Brett Griffin Page 13

14 Figure 3: Carpet Figure 4: Floor board Prepared by Brett Griffin Page 14

15 Figure 5: Bed sheet Figure 6: Ceramic tile Prepared by Brett Griffin Page 15

16 Figure 7: Circle showing the three segments Semen of each dilution was then pipetted into the corresponding marked circle on each of the substrates. These samples were locked and secured in an examination room contained within the Penrith Crime Scene Section for the duration of the experiments. Prepared by Brett Griffin Page 16

17 Products ABAcard p30 Test Kit Contains 25 Test Devices (in individually sealed pouches) 25 Sterile cotton swabs (in individually sealed packets) 25 Tubes containing extraction buffer 25 droppers (1 located in each test device pouch) Test Instructions (Annexure A) Figure 8: ABAcard p30 Test Kit box Figure 9: Contents of ABAcard p30 Test Kit box Prepared by Brett Griffin Page 17

18 Figures 10 and 11: Extraction buffer solution vial and p30 Test Device and dropper Figure 11: View of the ABAcard p30 Test Kit test device. Prepared by Brett Griffin Page 18

19 Preparation: (taken from enclosed instruction sheet contained within the kit) Preparation: For fresh stains/swabs: Extraction of specimens from fresh stains or fresh swab may be performed in the extraction buffer that is provided with this kit. You may fill in the relevant case identification information on the extraction tube label. Soak/extract the specimen/swab for at least 5 minutes at room temperature in the entire volume of the buffer. Gently mix/rotate the contents of the tube for at least 15 seconds with the swab. Press the swab tip gently against the tube wall to extract the liquid that is held in the swab. The extract from the specimen may now be used immediately for testing. Follow the Test Protocol. For aged/dried/wash stains: Follow the above preparation with an extraction time of 30 minutes at room temperature. Test Protocol 1. Ensure that the extract is at room temperature (warm it to room temperature if refrigerated) 2. Remove the device and the dropper from the sealed pouch. 3. Label the device with the case number. 4. Add 4-6 drops of sample (200µL) to the sample well S of the test device. 5. Read result at 10 minutes. Positive results can be seen as early as 1 minute depending upon the p30 concentration. For negative results, one must wait full 10 minutes. Positive Result 3 Negative Result 4 Invalid Result 5 3 POSITIVE. If there are two pink lines, one each in the test area "T" and in the control area "C", the test result is positive and indicates that the p30 level is at or above 4 ng / ml. 4 NEGATIVE. If there is only one pink line (in the control area "C"), the test result is Negative. This may indicate that (a) No p30 is present above 4ng/ml or (b) Presence of "High Dose Hook Effect". Presence of "High Dose Hook Effect" may give false negative result due to the presence of high concentration of p30 in the sample, as for example in undiluted seminal fluid. In such cases the sample may be retested using a 10 to 10,000 fold dilution. 5 INVALID. If there is no pink line visible in the control area "C", the test is inconclusive. Repeat the test and re-examine the test procedure carefully. Prepared by Brett Griffin Page 19

20 Phosphatesmo KM test strips (Directions directly from the Forensic Services Group Crime Scene Procedures Manual Specialist Manual) Figure 12: Phosphatesmo KM TM test strips Test a KM TM strip from the batch which is to be used with a known semen control prior to use Application 1) Check the expiry date on the Phosphatesmo KM strips. Do not use the strips if the expiry date has passed. 2) Expose the test paper by pulling apart the ends of the tape. Take care not to touch the test paper zone 3) Moisten the test paper with a drop of distilled water 4) Place the test paper directly on the semen sample, press firmly and wait 30 seconds 5) Examine the test paper for any colour development 6) Dispose of the used control as contaminated waste Prepared by Brett Griffin Page 20

21 Interpretation 1) A positive result was recorded when a deep violet colour develops within 30 seconds 2) Record a negative result if any colour other than violet develops 3) Record a negative result if any colouration develops after one minute 4) Take care to observe and record colour changes within 30 seconds of application as purple colouration may develop over time due to oxidation in air 5) If no colour develops within 30 seconds, the Phosphatesmo KM strips may be inactive due to exposure of the strips to heat or passing of the expiry date. Do not use strips whose expiry date has passed. 6) Alternatively, the semen control may have degraded significantly and may no longer contain acid phosphatase at a detectable level 7) If a negative result is obtained with the known semen control, return the Phosphatesmo KM strips to the scientific officer for further testing 8) Open a new pack of Phosphatesmo KM strips and conduct the known semen control test again 9) If a second negative result is obtained with the known semen control, it is likely that the known semen is degraded 10) Return the suspect semen control to the scientific officer for further testing and use a new semen control. Prepared by Brett Griffin Page 21

22 Application 1) Do not touch the test strip with your bare hands. 2) Due to the expense of the tests strips, cut each strip into 3 or 4 pieces just prior to use. 3) Once open and cut the test strips must be used immediately. Any unused pieces should be discarded at the completion of the examination, 4) Locate any suspected seminal stains by visual examination. 5) Taking care not to touch the test paper zone, expose the tests paper by pulling apart the ends of the tape. 6) Moisten the test paper with a drop of distilled water. 7) Place the test paper directly on the stain and press firmly. 8) Wait 30 seconds. 9) Examine the test paper for any colour development. 10) If the item/area is to be collected for further analysis, record, package, label and seal it. 11) If the item/area cannot be collected but further analysis is required, the identified area should be swabbed. Prepared by Brett Griffin Page 22

23 Interpretation of Findings A positive result is recorded when a deep violet colour develops within 30 seconds A negative result is recorded if any other colour than violet develops A negative result is recorded if any colouration develops after one minute Take care to observe and record colour changes within 30 seconds of application, as purple colouration may develop over time due to oxidation in air Prepared by Brett Griffin Page 23

24 Results Experiment 1 Aim - To determine the sensitivity limitations of the ABAcard p30 Test Kits, comparing my results against the manufacturers sensitivity claim of positive results of up to 1: dilutions. I used the following dilutions, Whole, 1:10, 1:100, 1:1000, 1:10000, 1: and 1: Method Dip the supplied cotton swab into the dilution, then the cotton tip was broken off into buffer solution and was allowed to remain there for 5 minutes. 6 drops of this buffer solution was then pipetted into the sample well of the ABAcard p30 Test Kit. I then waited 10 mins and recorded the result. This was replicated for all 7 dilutions. Results The ABAcard p30 Test Kit was able to detect up to the manufacturers claimed 1: , with a very weak reaction at that dilution. Figure 13: Results of ABAcard p30 Test Kits with liquid samples. Prepared by Brett Griffin Page 24

25 Figure 14: Close view of results of ABAcard p30 Test Kits with liquid semen. Semen Dilution Result of test - ABAcard Whole Semen S 1 : 10 S 1 : 100 S 1 : 1000 S 1 : M 1 : W 1 : VW Table 2: Results of the ABAcard p30 Test Kits and various semen dilutions. The results of all of these experiments were recorded using qualitative measures. The results were tabulated by dilution strength. The results were recorded using the following criteria: NR No reaction observed VW Very Weak reaction W Weak reaction M Medium reaction S Strong reaction Prepared by Brett Griffin Page 25

26 Experiment 2 Aim - To determine the sensitivity limitations of the Phosphatesmo KM TM strips, comparing my results against those of the ABAcard p30 Test Kit, using the same dilutions up to 1: Method A drop of the various dilutions was pipetted onto individual Phosphatesmo KM TM strips. I waited the recommended 30 secs and recorded my results. Results The Phosphatesmo KM TM strips gave a positive reaction only up to 1: With this in mind, the following experiments with Phosphatesmo KM TM strips were limited up to 1:10000 Figure 15: Phosphatesmo KM TM test strips results with liquid sample. Prepared by Brett Griffin Page 26

27 Figure 16: Close view of the Phosphatesmo KM TM results. Semen Dilution Result of test - Phosphatesmo KM TM Whole Semen S 1 : 10 S 1 : 100 S 1 : 1000 M 1 : W 1 : NR 1 : NR Table 3: Results of the Phosphatesmo KM TM test strips and various semen dilutions. NR No reaction observed VW Very Weak reaction W Weak reaction M Medium reaction S Strong reaction Prepared by Brett Griffin Page 27

28 Experiment 3 Aim - To determine the ABAcard p30 Test Kits ability to detect stains of various dilutions (Whole, 1:10, 1:100, 1:1000, 1:10000, 1: and 1: ) deposited onto Carpet. Test carried out 3 days following deposition. Method - Dip the supplied cotton swab into the buffer solution to hydrate the swab tip. I then rubbed the cotton swab tip in the target area. This cotton swab tip was broken off into buffer solution and was allowed to remain there for 5 minutes. 6 drops of this buffer solution was then pipetted into the sample well of the ABAcard p30 Test Kit. I then waited 30 mins and recorded the result. This was replicated for all 7 dilutions. Results The ABAcard p30 Test Kit was able to give a positive result for all dilutions, with a very weak reaction at 1: Figure 17: ABAcard p30 Test Kit Carpet Day 3 Prepared by Brett Griffin Page 28

29 Figure 18: Close view of the ABAcard p30 Test Kits Carpet Day 3 Semen Dilution Result of test ABAcard Carpet 3 Days Whole Semen S 1 : 10 S 1 : 100 S 1 : 1000 S 1 : W 1 : VW 1 : VW Table 4: Results of the ABAcard p30 Test Kits and various semen dilutions on carpet at day 3. NR No reaction observed VW Very Weak reaction W Weak reaction M Medium reaction S Strong reaction Prepared by Brett Griffin Page 29

30 Experiment 4 Aim - To determine the Phosphatesmo KM TM strips ability to detect stains of various dilutions (Whole, 1:10, 1:100, 1:1000, 1:10000) deposited onto Carpet. Test carried out 3 days following deposition. Method - Place a single drop of sterile water onto the test area of the Phosphatesmo KM TM strip. This test area was then pressed against the stain for the recommended 30 secs and I then recorded my results. This was replicated for dilutions, Whole, 1:10, 1:100, 1:1000, 1:10000) Results - Phosphatesmo KM TM strips returned positive result for all dilutions, with a very weak reaction at 1: Figure 19: Phosphatesmo KM TM Carpet Day 3 Prepared by Brett Griffin Page 30

31 Figure 20: Results of Phosphatesmo KM TM Carpet Day 3 Semen Dilution Result of test Phosphatesmo KM TM Carpet 3 Days Whole Semen S 1 : 10 S 1 : 100 S 1 : 1000 M 1 : W Table 5: Results of the Phosphatesmo KM TM test strips and various semen dilutions on carpet at day 3. NR No reaction observed VW Very Weak reaction W Weak reaction M Medium reaction S Strong reaction Prepared by Brett Griffin Page 31

32 Experiment 5 Aim - To determine ABAcard p30 Test Kits ability to detect stains of various dilutions (Whole, 1:10, 1:100, 1:1000, 1:10000, 1: and 1: ) deposited onto a floorboard. Test carried out 3 days following deposition. Method - Dip the supplied cotton swab into the buffer solution to hydrate the swab tip. I then rubbed the cotton swab tip in the target area. This cotton swab tip was broken off into buffer solution and was allowed to remain there for 5 minutes. 6 drops of this buffer solution was then pipetted into the sample well of the ABAcard p30 Test Kit. I then waited 30 mins and recorded the result. This was replicated for all 7 dilutions. Results The ABAcard p30 Test Kit was able to give a positive result for all dilutions. Figure 21: ABAcard p30 Test Kits board Day 3 Prepared by Brett Griffin Page 32

33 Figure 22: Close view of the ABAcard p30 Test Kits Floorboard Day 3 Semen Dilution Result of test ABAcard Floor board 3 Days Whole Semen S 1 : 10 S 1 : 100 S 1 : 1000 M 1 : W 1 : W 1 : W Table 6: Results of the ABAcard p30 Test Kits and various semen dilutions on floorboards at day 3. NR No reaction observed VW Very Weak reaction W Weak reaction M Medium reaction S Strong reaction Prepared by Brett Griffin Page 33

34 Experiment 6 Aim - To determine the Phosphatesmo KM TM strips ability to detect stains of various dilutions (Whole, 1:10, 1:100, 1:1000, 1:10000) deposited onto a floorboard. Test carried out 3 days following deposition. Method - Place a single drop of sterile water onto the test area of the Phosphatesmo KM TM strip. This test area was then pressed against the stain for the recommended 30 secs and I then recorded my results. This was replicated for dilutions, Whole, 1:10, 1:100, 1:1000, 1:10000) Results - Phosphatesmo KM TM strips returned positive results through all dilutions however 1:10000 was very weak. Figure 23: Phosphatesmo KM TM test strips Floor board Day 3 Prepared by Brett Griffin Page 34

35 Figure 24: Close view Phosphatesmo KM TM test strips Floor board Day 3 Semen Dilution Result of test Phosphatesmo KM TM Floor board 3 Days Whole Semen S 1 : 10 S 1 : 100 S 1 : 1000 M 1 : W Table 7: Results of the Phosphatesmo KM TM test strips and various semen dilutions on floorboards at day 3. NR No reaction observed VW Very Weak reaction W Weak reaction M Medium reaction S Strong reaction Prepared by Brett Griffin Page 35

36 Experiment 7 Aim - To determine the ABAcard p30 Test Kits ability to detect stains of various dilutions (Whole, 1:10, 1:100, 1:1000, 1:10000, 1: and 1: ) deposited onto a bed sheet. Test carried out 3 days following deposition. Method - Dip the supplied cotton swab into the buffer solution to hydrate the swab tip. I then rubbed the cotton swab tip in the target area. This cotton swab tip was broken off into buffer solution and was allowed to remain there for 5 minutes. 6 drops of this buffer solution was then pipetted into the sample well of the ABAcard p30 Test Kit. I then waited 30 mins and recorded the result. This was replicated for all 7 dilutions. Results The ABAcard p30 Test Kit was able to give a positive result up to 1:1000. Figure 25: ABAcard p30 Test Kits Bedsheet Day 3 Prepared by Brett Griffin Page 36

37 Figure 26: Close view of the ABAcard p30 Test Kits Bedsheet Day 3 Semen Dilution Result of test ABAcard Bed sheet 3 Days Whole Semen S 1 : 10 S 1 : 100 M 1 : 1000 VW 1 : NR 1 : NR 1 : NR Table 8: Results of the ABAcard p30 Test Kits and various semen dilutions on a bedsheet at day 3. NR No reaction observed VW Very Weak reaction W Weak reaction M Medium reaction S Strong reaction Prepared by Brett Griffin Page 37

38 Experiment 8 Aim - To determine the Phosphatesmo KM TM strips ability to detect stains of various dilutions (Whole, 1:10, 1:100, 1:1000, 1:10000) deposited onto a bed sheet. Test carried out 3 days following deposition. Method - Place a single drop of sterile water onto the test area of the Phosphatesmo KM TM strip. This test area was then pressed against the stain for the recommended 30 secs and I then recorded my results. This was replicated for dilutions, Whole, 1:10, 1:100, 1:1000, 1:10000) Results - Phosphatesmo KM TM strips returned positive results up to 1:1000. Figure 27: Phosphatesmo KM TM test strips Bedsheet Day 3 Prepared by Brett Griffin Page 38

39 Figure 28: Close view Phosphatesmo KM TM test strips Bedsheet Day 3 Semen Dilution Result of test Phosphatesmo KM TM Bed sheet 3 Days Whole Semen S 1 : 10 S 1 : 100 M 1 : 1000 VW 1 : NR Table 9: Results of the Phosphatesmo KM TM test strips and various semen dilutions on a bedsheet at day 3. NR No reaction observed VW Very Weak reaction W Weak reaction M Medium reaction S Strong reaction Prepared by Brett Griffin Page 39

40 Experiment 9 Aim - To determine the ABAcard p30 Test Kits ability to detect stains of various dilutions (Whole, 1:10, 1:100, 1:1000, 1:10000, 1: and 1: ) deposited onto a tile. Test carried out 3 days following deposition. Method - Dip the supplied cotton swab into the buffer solution to hydrate the swab tip. I then rubbed the cotton swab tip in the target area. This cotton swab tip was broken off into buffer solution and was allowed to remain there for 5 minutes. 6 drops of this buffer solution was then pipetted into the sample well of the ABAcard p30 Test Kit. I then waited 30 mins and recorded the result. This was replicated for all 7 dilutions. Results The ABAcard p30 Test Kit was able to give a positive result up to 1:1000. Figure 29: ABAcard p30 Test Kits Tile Day 3 Prepared by Brett Griffin Page 40

41 Figure 30: Close view of the ABAcard p30 Test Kits Tile Day 3 Semen Dilution Result of test ABAcard Tile - 3 Days Whole Semen S 1 : 10 S 1 : 100 M 1 : 1000 VW 1 : NR 1 : NR 1 : NR Table 10: Results of the ABAcard p30 Test Kits and various semen dilutions on a tile at day 3. NR No reaction observed VW Very Weak reaction W Weak reaction M Medium reaction S Strong reaction Prepared by Brett Griffin Page 41

42 Experiment 10 Aim - To determine the Phosphatesmo KM TM strips ability to detect stains of various dilutions (Whole, 1:10, 1:100, 1:1000, 1:10000) deposited onto a tile. Test carried out 3 days following deposition. Method - Place a single drop of sterile water onto the test area of the Phosphatesmo KM TM strip. This test area was then pressed against the stain for the recommended 30 secs and I then recorded my results. This was replicated for dilutions, Whole, 1:10, 1:100, 1:1000, 1:10000) Results - Phosphatesmo KM TM strips returned positive results returned positive results through all dilutions. Figure 31: Phosphatesmo KM TM test strips Tile Day 3 Prepared by Brett Griffin Page 42

43 Figure 32: Close view Phosphatesmo KM TM test strips Tile Day 3 Semen Dilution Result of test Phosphatesmo KM TM Tile - 3 Days Whole Semen S 1 : 10 S 1 : 100 S 1 : 1000 M 1 : VW Table 11: Results of the Phosphatesmo KM TM test strips and various semen dilutions on a tile at day 3 NR No reaction observed VW Very Weak reaction W Weak reaction M Medium reaction S Strong reaction Prepared by Brett Griffin Page 43

44 Experiment 11 Aim - To determine the ABAcard p30 Test Kits ability to detect stains of various dilutions (Whole, 1:10, 1:100, 1:1000, 1:10000, 1: and 1: ) deposited onto Carpet. Test carried out 2 weeks following deposition. Method - Dip the supplied cotton swab into the buffer solution to hydrate the swab tip. I then rubbed the cotton swab tip in the target area. This cotton swab tip was broken off into buffer solution and was allowed to remain there for 5 minutes. 6 drops of this buffer solution was then pipetted into the sample well of the ABAcard p30 Test Kit. I then waited 30 mins and recorded the result. This was replicated for all 7 dilutions. Results The ABAcard p30 Test Kit returned positive results through all dilutions. Figure 33: ABAcard p30 Test Kits Carpet 2 weeks Prepared by Brett Griffin Page 44

45 Figure 34: Close view of the ABAcard p30 Test Kits Carpet 2 weeks Semen Dilution Result of test ABAcard Carpet 2 weeks Whole Semen S 1 : 10 S 1 : 100 S 1 : 1000 M 1 : VW 1 : VW 1 : NR Table 12: Results of the ABAcard p30 Test Kits and various semen dilutions on carpet at 2 weeks. NR No reaction observed VW Very Weak reaction W Weak reaction M Medium reaction S Strong reaction Prepared by Brett Griffin Page 45

46 Experiment 12 Aim - To determine the Phosphatesmo KM TM strips ability to detect stains of various dilutions (Whole, 1:10, 1:100, 1:1000, 1:10000) deposited onto Carpet. Test carried out 2 weeks following deposition. Method - Place a single drop of sterile water onto the test area of the Phosphatesmo KM TM strip. This test area was then pressed against the stain for the recommended 30 secs and I then recorded my results. This was replicated for dilutions, Whole, 1:10, 1:100, 1:1000, 1:10000) Results - Phosphatesmo KM TM strips returned positive results through all dilutions. Figure 35: Phosphatesmo KM TM test strips Carpet 2 weeks Prepared by Brett Griffin Page 46

47 Figure 36: Close view Phosphatesmo KM TM test strips Carpet 2 weeks Semen Dilution Result of test Phosphatesmo KM TM Carpet - 2 weeks Whole Semen S 1 : 10 S 1 : 100 M 1 : 1000 M 1 : W Table 13: Results of the Phosphatesmo KM TM test strips and various semen dilutions on carpet at 2 weeks. NR No reaction observed VW Very Weak reaction W Weak reaction M Medium reaction S Strong reaction Prepared by Brett Griffin Page 47

48 Experiment 13 Aim - To determine the ABAcard p30 Test Kits ability to detect stains of various dilutions (Whole, 1:10, 1:100, 1:1000, 1:10000, 1: and 1: ) deposited onto a floorboard. Test carried out 2 weeks following deposition. Method - Dip the supplied cotton swab into the buffer solution to hydrate the swab tip. I then rubbed the cotton swab tip in the target area. This cotton swab tip was broken off into buffer solution and was allowed to remain there for 5 minutes. 6 drops of this buffer solution was then pipetted into the sample well of the ABAcard p30 Test Kit. I then waited 30 mins and recorded the result. This was replicated for all 7 dilutions. Results The ABAcard p30 Test Kit was able to give a positive up to 1:10000 (strip not visible in photograph however was very faint) Figure 37: ABAcard p30 Test Kits Floorboard 2 weeks Prepared by Brett Griffin Page 48

49 Figure 38: Close view of the ABAcard p30 Test Kits Floorboard 2 weeks Semen Dilution Result of test ABAcard Floor board 2 weeks Whole Semen S 1 : 10 S 1 : 100 S 1 : 1000 W 1 : VW 1 : NR 1 : NR Table 14: Results of the ABAcard p30 Test Kits and various semen dilutions on a floorboard at 2 weeks. NR No reaction observed VW Very Weak reaction W Weak reaction M Medium reaction S Strong reaction Prepared by Brett Griffin Page 49

50 Experiment 14 Aim - To determine the Phosphatesmo KM TM strips ability to detect stains of various dilutions (Whole, 1:10, 1:100, 1:1000, 1:10000) deposited onto a floorboard. Test carried out 2 weeks following deposition. Method - Place a single drop of sterile water onto the test area of the Phosphatesmo KM TM strip. This test area was then pressed against the stain for the recommended 30 secs and I then recorded my results. This was replicated for dilutions, Whole, 1:10, 1:100, 1:1000, 1:10000) Results - Phosphatesmo KM TM strips returned positive results through all dilutions. Figure 39: Phosphatesmo KM TM test strips Floorboard 2 weeks Prepared by Brett Griffin Page 50

51 Figure 40: Close view Phosphatesmo KM TM test strips Floorboard 2 weeks Semen Dilution Result of test Phosphatesmo KM TM Floor board - 2 weeks Whole Semen 1 : 10 S 1 : 100 M 1 : 1000 W 1 : W Table 15: Results of the Phosphatesmo KM TM test strips and various semen dilutions on a floorboard at 2 weeks. S NR No reaction observed VW Very Weak reaction W Weak reaction M Medium reaction S Strong reaction Prepared by Brett Griffin Page 51

52 Experiment 15 Aim - To determine the ABAcard p30 Test Kits ability to detect stains of various dilutions (Whole, 1:10, 1:100, 1:1000, 1:10000, 1: and 1: ) deposited onto a bed sheet. Test carried out 2 weeks following deposition. Method - Dip the supplied cotton swab into the buffer solution to hydrate the swab tip. I then rubbed the cotton swab tip in the target area. This cotton swab tip was broken off into buffer solution and was allowed to remain there for 5 minutes. 6 drops of this buffer solution was then pipetted into the sample well of the ABAcard p30 Test Kit. I then waited 30 mins and recorded the result. This was replicated for all 7 dilutions. Results The ABAcard p30 Test Kit returned positive results through all dilutions. Figure 41: ABAcard p30 Test Kits Bed sheet 2 weeks Prepared by Brett Griffin Page 52

53 Figure 42: Close view of the ABAcard p30 Test Kits Bed sheet 2 weeks Semen Dilution Result of test ABAcard Bed sheet 2 weeks Whole Semen S 1 : 10 S 1 : 100 M 1 : 1000 M 1 : W 1 : VW 1 : VW Table 16: Results of the ABAcard p30 Test Kits and various semen dilutions on a bedsheet at 2 weeks. NR No reaction observed VW Very Weak reaction W Weak reaction M Medium reaction S Strong reaction Prepared by Brett Griffin Page 53

54 Experiment 16 Aim - To determine the Phosphatesmo KM TM strips ability to detect stains of various dilutions (Whole, 1:10, 1:100, 1:1000, 1:10000) deposited onto a bed sheet. Test carried out 2 weeks following deposition. Method - Place a single drop of sterile water onto the test area of the Phosphatesmo KM TM strip. This test area was then pressed against the stain for the recommended 30 secs and I then recorded my results. This was replicated for dilutions, Whole, 1:10, 1:100, 1:1000, 1:10000) Results - Phosphatesmo KM TM strips returned positive results up to 1:1000. Figure 43: Close view Phosphatesmo KM TM test strips Bedsheet 2 weeks Prepared by Brett Griffin Page 54

55 Figure 44: Close view Phosphatesmo KM TM test strips Bedsheet 2 weeks Semen Dilution Result of test Phosphatesmo KM TM Bed sheet - 2 weeks Whole Semen S 1 : 10 S 1 : 100 W 1 : 1000 VW 1 : NR Table 17: Results of the Phosphatesmo KM TM test strips and various semen dilutions on a bedsheet at 2 weeks. NR No reaction observed VW Very Weak reaction W Weak reaction M Medium reaction S Strong reaction Prepared by Brett Griffin Page 55

56 Experiment 17 Aim - To determine the ABA cards ability to detect stains of various dilutions (Whole, 1:10, 1:100, 1:1000, 1:10000, 1: and 1: ) deposited onto a tile. Test carried out 2 weeks following deposition. Method - Dip the supplied cotton swab into the buffer solution to hydrate the swab tip. I then rubbed the cotton swab tip in the target area. This cotton swab tip was broken off into buffer solution and was allowed to remain there for 5 minutes. 6 drops of this buffer solution was then pipetted into the sample well of the ABAcard p30 Test Kits. I then waited 30 mins and recorded the result. This was replicated for all 7 dilutions. Results The ABA card was able to give a positive result up to 1: Figure 45: ABAcard p30 Test Kits Tile 2 Weeks Prepared by Brett Griffin Page 56

57 Figure 46: Close view of the ABAcard p30 Test Kits Tile 2 Weeks Semen Dilution Result of test ABAcard Tile 2 weeks Whole Semen S 1 : 10 S 1 : 100 S 1 : 1000 W 1 : VW 1 : NR 1 : NR Table 18: Results of the ABAcard p30 Test Kits and various semen dilutions on a tile at 2 weeks. NR No reaction observed VW Very Weak reaction W Weak reaction M Medium reaction S Strong reaction Prepared by Brett Griffin Page 57

58 Experiment 18 Aim - To determine the Phosphatesmo KM TM strips ability to detect stains of various dilutions (Whole, 1:10, 1:100, 1:1000, 1:10000) deposited onto a tile. Test carried out 2 weeks following deposition. Method - Place a single drop of sterile water onto the test area of the Phosphatesmo KM TM strip. This test area was then pressed against the stain for the recommended 30 secs and I then recorded my results. This was replicated for dilutions, Whole, 1:10, 1:100, 1:1000, 1:10000) Results - Phosphatesmo KM TM strips returned positive results through all dilutions. Figure 47: Phosphatesmo KM TM test strips Tile 2 weeks Prepared by Brett Griffin Page 58

59 Figure 48: Close view Phosphatesmo KM TM test strips Tile 2 weeks Semen Dilution Result of test Phosphatesmo KM TM Tile - 2 weeks Whole Semen S 1 : 10 S 1 : 100 S 1 : 1000 M 1 : M Table 19: Results of the Phosphatesmo KM TM test strips and various semen dilutions on a tile at 2 weeks. NR No reaction observed VW Very Weak reaction W Weak reaction M Medium reaction S Strong reaction Prepared by Brett Griffin Page 59

60 Experiment 19 Aim - To determine the ABAcard p30 Test Kits ability to detect stains of various dilutions (Whole, 1:10, 1:100, 1:1000, 1:10000, 1: and 1: ) deposited onto Carpet. Test carried out 4 weeks following deposition. Method - Dip the supplied cotton swab into the buffer solution to hydrate the swab tip. I then rubbed the cotton swab tip in the target area. This cotton swab tip was broken off into buffer solution and was allowed to remain there for 5 minutes. 6 drops of this buffer solution was then pipetted into the sample well of the ABAcard p30 Test Kit. I then waited 30 mins and recorded the result. This was replicated for all 7 dilutions. Results The ABAcard p30 Test Kit returned positive results through all dilutions. Figure 49: ABAcard p30 Test Kits Carpet 4 weeks Prepared by Brett Griffin Page 60

61 Figure 50: Close view of the ABAcard p30 Test Kits Carpet Day 4 weeks Semen Dilution Result of test ABAcard Carpet 4 weeks Whole Semen S 1 : 10 S 1 : 100 S 1 : 1000 S 1 : M 1 : W 1 : VW Table 20: Results of the ABAcard p30 Test Kits and various semen dilutions on carpet at 4 weeks. NR No reaction observed VW Very Weak reaction W Weak reaction M Medium reaction S Strong reaction Prepared by Brett Griffin Page 61

62 Experiment 20 Aim - To determine the Phosphatesmo KM TM strips ability to detect stains of various dilutions (Whole, 1:10, 1:100, 1:1000, 1:10000) deposited onto Carpet. Test carried out 4 weeks following deposition. Method - Place a single drop of sterile water onto the test area of the Phosphatesmo KM TM strip. This test area was then pressed against the stain for the recommended 30 secs and I then recorded my results. This was replicated for dilutions, Whole, 1:10, 1:100, 1:1000, 1:10000) Results - Phosphatesmo KM TM strips returned positive results through all dilutions, however 1:10000 was very weak. Figure 51: Phosphatesmo KM TM test strips Carpet 4 weeks Prepared by Brett Griffin Page 62

63 Figure 52: Close view Phosphatesmo KM TM test strips Carpet 4 weeks Semen Dilution Result of test Phosphatesmo KM TM Carpet - 4 weeks Whole Semen S 1 : 10 S 1 : 100 M 1 : 1000 VW 1 : VW Table 21: Results of the Phosphatesmo KM TM test strips and various semen dilutions on carpet at 4 weeks. NR No reaction observed VW Very Weak reaction W Weak reaction M Medium reaction S Strong reaction Prepared by Brett Griffin Page 63

64 Experiment 21 Aim - To determine the ABAcard p30 Test Kits ability to detect stains of various dilutions (Whole, 1:10, 1:100, 1:1000, 1:10000, 1: and 1: ) deposited onto a floorboard. Test carried out 4 weeks following deposition. Method - Dip the supplied cotton swab into the buffer solution to hydrate the swab tip. I then rubbed the cotton swab tip in the target area. This cotton swab tip was broken off into buffer solution and was allowed to remain there for 5 minutes. 6 drops of this buffer solution was then pipetted into the sample well of the ABAcard p30 Test Kit. I then waited 30 mins and recorded the result. This was replicated for all 7 dilutions. Results The ABAcard p30 Test returned positive results through all dilutions. Figure 53: Close view of the ABAcard p30 Test Kits Floor board 4 weeks Prepared by Brett Griffin Page 64

65 Figure 54: Close view of the ABAcard p30 Test Kits Floor board 4 weeks Semen Dilution Result of test ABAcard Floor board 4 weeks Whole Semen S 1 : 10 S 1 : 100 S 1 : 1000 M 1 : W 1 : VW 1 : VW Table 22: Results of the ABAcard p30 Test Kits and various semen dilutions on a floorboard at 4 weeks. NR No reaction observed VW Very Weak reaction W Weak reaction M Medium reaction S Strong reaction Prepared by Brett Griffin Page 65

66 Experiment 22 Aim - To determine the Phosphatesmo KM TM strips ability to detect stains of various dilutions (Whole, 1:10, 1:100, 1:1000, 1:10000) deposited onto a floorboard. Test carried out 4 weeks following deposition. Method - Place a single drop of sterile water onto the test area of the Phosphatesmo KM TM strip. This test area was then pressed against the stain for the recommended 30 secs and I then recorded my results. This was replicated for dilutions, Whole, 1:10, 1:100, 1:1000, 1:10000) Results - Phosphatesmo KM TM strips returned positive results through all dilutions. Figure 55: Phosphatesmo KM TM test strips Floorboard 4 weeks Prepared by Brett Griffin Page 66

67 Figure 56: Close view Phosphatesmo KM TM test strips Floorboard 4 weeks Semen Dilution Result of test Phosphatesmo KM TM Floor board - 4 weeks Whole Semen 1 : 10 S 1 : 100 M 1 : 1000 M 1 : W Table 23: Results of the Phosphatesmo KM TM test strips and various semen dilutions on a floorboard at 4 weeks. S NR No reaction observed VW Very Weak reaction W Weak reaction M Medium reaction S Strong reaction Prepared by Brett Griffin Page 67

68 Experiment 23 Aim - To determine the ABAcard p30 Test Kits ability to detect stains of various dilutions (Whole, 1:10, 1:100, 1:1000, 1:10000, 1: and 1: ) deposited onto a bed sheet. Test carried out 4 weeks following deposition. Method - Dip the supplied cotton swab into the buffer solution to hydrate the swab tip. I then rubbed the cotton swab tip in the target area. This cotton swab tip was broken off into buffer solution and was allowed to remain there for 5 minutes. 6 drops of this buffer solution was then pipetted into the sample well of the ABAcard p30 Test Kit. I then waited 30 mins and recorded the result. This was replicated for all 7 dilutions. Results The ABAcard p30 Test Kit returned positive results through all dilutions. Figure 57: Close view of the ABAcard p30 Test Kits Bedsheet 4 weeks Prepared by Brett Griffin Page 68

69 Figure 58: Close view of the ABAcard p30 Test Kits Bedsheet 4 weeks Semen Dilution Result of test ABAcard Bedsheet 4 weeks Whole Semen S 1 : 10 S 1 : 100 M 1 : 1000 M 1 : W 1 : W 1 : VW Table 24: Results of the ABAcard p30 Test Kits and various semen dilutions on a bedsheet at 4 weeks. NR No reaction observed VW Very Weak reaction W Weak reaction M Medium reaction S Strong reaction Prepared by Brett Griffin Page 69

70 Experiment 24 Aim - To determine the Phosphatesmo KM TM strips ability to detect stains of various dilutions (Whole, 1:10, 1:100, 1:1000, 1:10000) deposited onto a bed sheet. Test carried out 4 weeks following deposition. Method - Place a single drop of sterile water onto the test area of the Phosphatesmo KM TM strip. This test area was then pressed against the stain for the recommended 30 secs and I then recorded my results. This was replicated for dilutions, Whole, 1:10, 1:100, 1:1000, 1:10000) Results - Phosphatesmo KM TM strips returned positive results up to 1:100. Figure 59: Close view Phosphatesmo KM TM test strips Bedsheet 4 weeks Prepared by Brett Griffin Page 70

71 Figure 60: Close view Phosphatesmo KM TM test strips Bedsheet 4 weeks Semen Dilution Result of test Phosphatesmo KM TM Bedsheet - 4 weeks Whole Semen S 1 : 10 S 1 : 100 VW 1 : 1000 NR 1 : NR Table 25: Results of the Phosphatesmo KM TM test strips and various semen dilutions on a bedsheet at 4 weeks. NR No reaction observed VW Very Weak reaction W Weak reaction M Medium reaction S Strong reaction Prepared by Brett Griffin Page 71

72 Experiment 25 Aim - To determine the ABAcard p30 Test Kits ability to detect stains of various dilutions (Whole, 1:10, 1:100, 1:1000, 1:10000, 1: and 1: ) deposited onto a tile. Test carried out 4 weeks following deposition. Method - Dip the supplied cotton swab into the buffer solution to hydrate the swab tip. I then rubbed the cotton swab tip in the target area. This cotton swab tip was broken off into buffer solution and was allowed to remain there for 5 minutes. 6 drops of this buffer solution was then pipetted into the sample well of the ABAcard p30 Test Kit. I then waited 30 mins and recorded the result. This was replicated for all 7 dilutions. Results The ABAcard p30 Test Kit was able to give a positive result up to 1:1000. Figure 61: Close view of the ABAcard p30 Test Kits Tile 4 weeks Prepared by Brett Griffin Page 72

73 Figure 62: Close view of the ABAcard p30 Test Kits Tile 4 weeks Semen Dilution Result of test ABAcard Tile 4 weeks Whole Semen S 1 : 10 S 1 : 100 S 1 : 1000 W 1 : NR 1 : NR 1 : NR Table 26: Results of the ABAcard p30 Test Kits and various semen dilutions on a tile at 4 weeks. NR No reaction observed VW Very Weak reaction W Weak reaction M Medium reaction S Strong reaction Prepared by Brett Griffin Page 73

74 Experiment 26 Aim - To determine the Phosphatesmo KM TM strips ability to detect stains of various dilutions (Whole, 1:10, 1:100, 1:1000, 1:10000) deposited onto a tile. Test carried out 4 weeks following deposition. Method - Place a single drop of sterile water onto the test area of the Phosphatesmo KM TM strip. This test area was then pressed against the stain for the recommended 30 secs and I then recorded my results. This was replicated for dilutions, Whole, 1:10, 1:100, 1:1000, 1:10000) Results - Phosphatesmo KM TM strips returned positive results through all dilutions. Figure 63: Phosphatesmo KM TM test strips Tile 4 weeks Prepared by Brett Griffin Page 74

75 Figure 64: Close view Phosphatesmo KM TM test strips Tile 4 weeks Semen Dilution Result of test Phosphatesmo KM TM Bedsheet - 4 weeks Whole Semen S 1 : 10 S 1 : 100 M 1 : 1000 W 1 : W Table 27: Results of the Phosphatesmo KM TM test strips and various semen dilutions on a bedsheet at 4 weeks. NR No reaction observed VW Very Weak reaction W Weak reaction M Medium reaction S Strong reaction Prepared by Brett Griffin Page 75

76 Experiment 27 Aim - To determine ABAcard p30 Test Kits ability to still be effective after being exposed to high temperatures, as experienced inside a locked motor vehicle. Method I placed 1 x sealed kit into a work motor vehicle on Friday the 1 st October, 2010, and allowed it to remain within the vehicle for 1 week. I placed a second sealed kit into the same vehicle on Thursday the 7 th October, On Friday the 8 th October, 2010, I removed both sealed kits from the vehicle and returned them to the Penrith Crime Scene office. I then dipped 2 x supplied cotton swabs into the 1:10 dilution, then the cotton tips were broken off into the buffer solutions and was allowed to remain there for 5 minutes. 6 drops of these buffer solutions were then pipetted into the sample well of both of the ABAcard p30 Test Kits. I then waited 10 mins and recorded the result. Result Both ABAcard p30 Test Kits returned positive results at the 1:10 dilution after being exposed to high temperatures associated with locked vehicles.. Figure 65: ABAcard p30 Test Kits after being exposed to high temperatures in locked motor vehicle Prepared by Brett Griffin Page 76