Evaluation of mRNA-based approach for identification of saliva and semen

Sakurada, Koichi; Ikegaya, Hiroshi; Fukushima, Hisashi; Akutsu, Tomoko; Watanabe, Ken; Yoshino, Mineo

doi:10.1016/j.legalmed.2008.10.002

Cited by 86 publications

(47 citation statements)

References 11 publications

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“…These results suggest that humidity and light aŠect mRNA degradation as we previously reported. 15) As the relative Ct value of semen stains was signiˆcantly higher than that of saliva and blood stains at 4°C after 1 month (Fig. 3A C), mRNA stability appears to diŠer between body ‰uids, supporting our previousˆnding that mRNA stability in nasal stains was lower than in saliva stains.…”

Section: Discussionsupporting

confidence: 84%

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Identification of Body Fluid Stains Using Real-time RT-PCR: Discrimination Between Salivary, Nasal, and Vaginal Secretions

Sakurada

Akutsu

Watanabe

et al. 2013

Jpn. J. Forensic. Sci. Tech.

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In a criminal investigation, it is important to clarify the origin of the extracted DNA from available biological samples. In recent years, a new approach for the identiˆcation of body ‰uid stains has been demonstrated and involves a comparison of speciˆc mRNA expression levels. Here, we used real-time RT-PCR to examine the expression levels of STATH, HTN3, MUC4, and ESR1 genes from salivary, nasal, and vaginal secretions which are often thought to be di‹cult to discriminate between. STATH was expressed at high levels in all salivary (dCt＝1.27±3.18, n＝ 10) and all nasal secretions (dCt＝-0.99±3.94, n＝8), and at low levels in 3 of the 10 vaginal secretions (dCt＝16.34-17.47). HTN3 was expressed at high levels in all salivary secretions (dCt＝0.67±3.08). MUC4 was expressed at relatively high levels in all vaginal secretions (dCt＝4.97±2.36, n＝10), and at moderate levels in 5 of the 10 salivary secretions (dCt＝8.55-10.40) and in 4 of the 8 nasal secretions (dCt ＝6.56-8.23). ESR1 was expressed at relatively high levels in all vaginal secretions (dCt＝5.79±2.90), but no expression was found in salivary and nasal samples. These results indicate the possibility for speciˆc discrimination between various body ‰uids. To test the practicality of this, gene sensitivity and stability were examined in saliva, semen, and blood stains. Several genes, including ACTB, STATH, HTN3, PRM2, SEMG1, and HBB, were shown to be detectable even in small volume (0.1 mL) stains and were also expressed in 1-year-old stains. mRNA stability was dependent on favorable environmental factors, such as dryness and shading. In addition, the successful detection of target genes from mixed stains and simulated casework samples might promise the utility of this assay. The mRNA assay therefore appears to be a novel tool for body ‰uid identiˆcation from biological samples.

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Section: Discussionsupporting

confidence: 84%

“…All saliva samples expressed high levels of STATH and HTN3, as previously reported. 15,16) and did not express ESR1 ( Table 1). All nasal samples also expressed high levels of STATH but not HTN3, as previously reported, 16) and did not express ESR1 ( Table 2).…”

Section: Discussionmentioning

confidence: 99%

Identification of Body Fluid Stains Using Real-time RT-PCR: Discrimination Between Salivary, Nasal, and Vaginal Secretions

Sakurada

Akutsu

Watanabe

et al. 2013

Jpn. J. Forensic. Sci. Tech.

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“…Statherin has been identified as a potent inhibitor of calcium phosphate precipitation [8,9]. Recently, it was reported that statherin mRNA would be useful as a marker for mRNA-based saliva identification because of its high specificity for saliva [10][11][12][13]. Besides, enzyme-linked immunosorbent assay (ELISA) procedures have been developed for identification of body fluids [14][15][16][17][18] as well as ABO blood grouping [19][20][21][22] in various forensic samples with an advantage of its applicability for crude samples without purification.…”

Section: Introductionmentioning

confidence: 99%

Applicability of ELISA detection of statherin for forensic identification of saliva

et al. 2009

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Statherin is a low molecular-weight phosphoprotein secreted from the parotid gland. Statherin mRNA was previously reported to be a useful marker for mRNA-based saliva identification. In this study, applicability of ELISA detection of statherin for forensic identification of saliva was investigated. The specificity and sensitivity of ELISA for detection of statherin were compared with those of ELISA for α-amylase and the Phadebas® amylase test. Statherin was specifically detected in saliva but not in other body fluids. In addition, statherin was successfully detected in aged saliva stains, mixed body fluids-saliva stains, and simulated casework samples. On the other hand, although ELISA for α-amylase showed higher sensitivity than ELISA for statherin, it was not specific enough to identify saliva. The Phadebas® amylase test also showed positive results in other body fluids that are known to have α-amylase activity; however, it is easy to use for screening forensic casework samples. In conclusion, ELISA for detection of statherin developed in this study could be an effective tool for the forensic identification of saliva because of its specificity for saliva among other body fluids. Forensic casework samples should be tested by ELISA detection or mRNA-based analysis for statherin, depending on the condition of the sample, to supplement presumptive tests for α-amylase, such as the Phadebas® amylase test.

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“…These studies should present countermeasures against these inhibitors, such as methods to purify samples, methods of applying resistant testing, and methods for alternative testing. Several alternative approaches (e.g., enzyme-linked immunosorbent assay, PCR, mass spectrometry) have been studied for identifying the bodily fluids used in forensics [8,[11][12][13][14]; however, these methods are not widely used in casework because they are not as convenient or cost effective as conventional presumptive tests. The key point is that when dealing with samples that are suspected to be contaminated and inhibited, the samples should be evaluated using multiple tests when possible.…”

Section: Discussionmentioning

confidence: 99%

Practical Issues of Inhibition in Forensic Body Fluid Identifications

Ohta¹

2017

FSAR

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Evaluation of mRNA-based approach for identification of saliva and semen

Cited by 86 publications

References 11 publications

Identification of Body Fluid Stains Using Real-time RT-PCR: Discrimination Between Salivary, Nasal, and Vaginal Secretions

Identification of Body Fluid Stains Using Real-time RT-PCR: Discrimination Between Salivary, Nasal, and Vaginal Secretions

Applicability of ELISA detection of statherin for forensic identification of saliva

Practical Issues of Inhibition in Forensic Body Fluid Identifications

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