100:. The Takayama reagent is added to a slide with a presumptive blood sample. The slide is dried at 115 degrees Celsius following the addition of the Takayama reagent. Then it is placed under a microscope and a positive result is the visualization of dark red, feathery crystals. For the immunochromatographic test, it functions similar to a pregnancy test where antigens present in blood are detected and a positive result is a band at the test site and control site. After performing the various tests an analyst can confirm the presence of human blood and continue to develop a
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tissue types should have unique proteins and mRNA based on their role in the body. MiRNAs are also an ideal target for forensic analysis because they are small compared to other cellular components, so they tend to resist degradation better than other tissue markers, which is important considering that case work samples are not always going to be in pristine condition. Finally, miRNAs have the potential to be co-extracted and analyzed at the same time as DNA, combining the two processes into one for biological sample analysis, saving time and sample.
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perform the confirmatory test. This can be limiting if the forensic sample being tested is small to begin with. Also, serology is often done before any downstream analyses like DNA, so if sample is limited in size to begin with performing serological analyses and obtaining a DNA profile successfully may not be possible. Currently, researchers are looking into ways to identify bodily fluids with more success and less sample needed, and an emerging way to do this is with micro RNAs.
140:. In order to initially detect semen, an alternative light source (ALS) is used. Under UV light, semen fluoresces making it visible to investigators to collect samples from a crime scene. A common presumptive test for detecting semen is called the acid phosphatase (AP) test. The AP test detects the enzyme acid phosphatase that is secreted from the prostate gland. However, this test is only presumptive because
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the loop-mediated isothermal amplification make identification of different body fluids. Using LAMP has reduced the time needed to get results, which is what the ultimate goal was. Although it has proven to decrease total and hands-on time needed to get a result, there are still kinks to work out before this method is used in many or all forensic labs.
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one drop of hydrogen peroxide. A positive result induces a color change to pink. Similar to the Kastle-Meyer test, a hemastix is also a catalytic test simplified to a specialized strip where the blood sample is extracted by a wet swab and placed directly on the hemastix. A positive result induces a colour change from yellow to dark green.
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gel, the saliva sample is added and allowed to diffuse through the gel overnight. Visualization is accomplished by adding iodine to the gel which stains the starch in the gel blue. If saliva is present, then the alpha-amylase breaks down the starch, creating a clear coloured circle around where the sample was placed.
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mid-piece and tail stain green. However, not all males release sperm in their semen. If a male is aspermic or oligospermic, they either have no sperm or a low sperm count. Vasectomized males will not release sperm either. When sperm cells are not present, a second confirmatory test, the p30/PSA test, is performed.
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Like the technique of extracting miRNA, researchers have been able to test one or more samples by extracting DNA and testing it in an instrument that most labs have readily available. This method has proven to produce more DNA than PCR based amplification. Researchers have also added other factors to
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that are used to regulate gene expression by either regulating translation (protein synthesis) or marking messenger RNA (mRNA) for degradation. Given their regulatory role, the theory is that different miRNAs would be present in different amounts in certain fluid or tissue types because each of those
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A presumptive test to detect saliva is the alpha-amylase test also known as the
Phadebas Test. This detection technique is based on the activity of the enzyme alpha-amylase which breaks down starches from food into smaller oligosaccharide molecules, starting digestion in the mouth. Using a petri dish
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to the slide and stained with nuclear fast red for 15 minutes, then rinsed with deionized water. Next, a green stain is applied for 10 seconds, then rinsed with ethanol. The slide is placed under a compound light microscope for sperm observation. If sperm are present, the heads will stain red and the
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in blood that transports oxygen and carbon dioxide. A sterile cotton swab is soaked in distilled water and applied to the area of suspected blood to pick up some of the sample. One drop of alcohol is applied to the swab, followed by the addition of one drop of the phenolphthalein reagent, followed by
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Current potential miRNA biomarkers: Research is still needed in order to narrow down potential biomarkers, as some tissues and fluids have the same miRNA expressed in different concentrations. To date, blood and semen miRNAs have been the most studied and have found promising candidate biomarkers.
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process in which a chemical is sprayed onto a surface where blood is suspected to be. The chemical reacts with traces of blood, producing a chemi-luminescence, or apparent glow, as a result of the chemical reaction that occurs. As with all presumptive tests, this technique can produce false positive
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and then separated/visualized using a capillary electrophoresis protocol. cDNA specific primers would have to be designed for your miRNA targets. The final output is an electropherogram that contains not only the STR profile of the sample, but also a peak representing which miRNA is present in that
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miRNA can be extracted using a number of commercially available kits, such as the Solid Phase QIAmp DNA mini kit. Ideally, like the QIAmp kit, the extraction method used is able to extract DNA and miRNA simultaneously, minimizing the amount of reactions and the amount of sample used. miRNAs can be
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Testing for different body fluids with traditional serological techniques, such as those listed above, is possible, but not without some drawbacks. Firstly, not all body fluids have a reliable confirmatory test, and those that do typically require a larger amount of the suspected stain in order to
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that is produced by the prostatic gland in males. The p30/PSA test is an immunochromatographic test that detects the presence of the antigen p30 in semen samples. This test functions similar to a pregnancy test, where if the antigen p30 is present a band will appear at the test site and a control
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The presumptive detection of urine can be done by alternative light sources or a paradimethylaminocinnamaldehyde test (DMAC). The DMAC will react with urea, uric acid or ammonia which can all be found in urine. When a sample with potential urine is found, 0.1% DMAC can be applied. If there is a
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positive reaction, a pink/magenta colour will be present on the stain. There are only presumptive tests for urine detection because the tests used target material that can be found in other bodily fluids. This can cause a lot of false positives and inaccurate results.
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is found in other bodily fluids. To perform the test, a drop of the reagent sodium alpha-napthyphosphate is added to the presumptive stain followed by a drop of fast blue B. A positive result of this test is a color change to dark purple.
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For confirmatory tests, there has not been as much research done compared to blood and semen. Since these tests specifically target amylase, confirmatory tests can not be done considering amylase can be found in other bodily fluids.
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band will appear to confirm if the test is working properly. If the confirmatory test is positive, then semen is present in the sample. From there an analyst could continue to develop a DNA profile with downstream applications.
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Tong D, Jin Y, Xue T, Ma X, Zhang J, Ou X, et al. (2015) Investigation of the
Application of miR10b and miR135b in the Identification of Semen Stains. PLoS ONE 10(9): e0137067. doi:10.1371/ journal.pone.0137067
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Hanson, E.K., Lubenow, H., Ballantyne, J. (2009). Identification of forensically relevant body fluids using a panel of differentially expressed microRNAs. Analytical
Biochemistry.387, 303-314.
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quantified using quantitative Real Time PCR, similar to traditional DNA samples. However, primers and probes would have to be designed for the miRNA targets in order to do so. Unlike routine
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Meer, D., Uchimoto, M., Williams,G.(2013). Simultaneous analysis of micro-RNA and DNA for determining the body fluid origin of DNA profiles. Journal of
Forensic Sciences. 58,4; 967-971
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Confirmatory tests for semen include the christmas tree stain and the p30/PSA RSID kit. For the christmas tree stain, the sample is extracted with sterile water in order to make a
220:, miRNA amplification requires an extra step before the PCR process. miRNA requires a reverse transcription step to convert the miRNA fragments into their complementary DNA (
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which gives the analyst an indication that a specific bodily fluid may be present, but cannot completely confirm its presence. Following the presumptive tests,
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test are typically used. The
Takayama Crystal Assay, which forms a ferro protoporphyrin ring by a reaction between pyridine and the iron atom of the
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Fundamentals of
Forensic Science By Max M. Houck, Jay A. Siegel p. 229 Publisher: Academic Press; 2 edition (February 3, 2010) Language: English
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tests have also been made in order to detect alpha-amylase, but they are not always reliable because there can be a lot of false positives.
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Criminal
Investigation by Ronald F. Becker P. 8 Publisher: Jones & Bartlett Publishers; 3 edition (August 22, 2008) Language: English
71:). To detect blood at a crime scene or in the laboratory, an array of tests can be used. The most publicized test by crime shows is the
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results due to metals and strong chemicals, such as bleach, that will also react. Another common presumptive test is the
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Courts, C., Madea, B. (2010). Micro-rna- a potential for forensic science. Forensic
Science International. 203; 106-111
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Blood is composed of liquid plasma and serum with solid components consisting of red blood cells (
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Ong, Sandy Y.; Wain, Adrian; Groombridge, Linda; Grimes, Eileen (June 2012).
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Peyrot des
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547:"Sourcebook in Forensic Serology, Immunology, and Biochemistry"
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For confirmatory tests, the
Takayama Crystal Assay or an
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Current research: Loop-mediated isothermal amplification
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635:"Salivary Amylase: Digestion and Metabolic Syndrome"
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116:(CE), etc., followed by profile interpretation.
84:test. This is a catalytic test that detects the
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152:on a microscope slide. The sample is then
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591:Kohlmeier, Amanda; Klock, Susan (2018),
517:"Search Results | Forensic Supplies, m7"
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692:"Amylase: Phadebas test, saliva"
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545:Gaensslen, R.E. (August 1983).
43:. Serology testing begins with
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123:Stained sperm under microscope
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690:Mullen, Carrie (2012-03-15),
757:10.1016/j.scijus.2011.07.007
597:Encyclopedia of Reproduction
490:"Science Fair Project Ideas"
242:Potential Biomarkers for ID
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136:from a male's penis due to
41:bloodstain pattern analysis
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554:U.S. Department of Justice
195:Current research: microRNA
651:10.1007/s11892-016-0794-7
162:prostate-specific antigen
114:Capillary Electrophoresis
110:Polymerase Chain Reaction
639:Current Diabetes Reports
160:PSA(p30) is known as a
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63:), white blood cells (
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428:Butler, John (2005).
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94:immunochromatographic
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432:Forensic DNA Typing
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469:Biology LibreTexts
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330:References
154:heat-fixed
134:ejaculated
98:heme group
86:heme group
65:leukocytes
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150:wet mount
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319:microRNA
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247:Blood
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761:PMID
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359:ISBN
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304:RSID
222:cDNA
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226:PCR
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Text is available under the Creative Commons Attribution-ShareAlike License. Additional terms may apply.
