Selective culturing and genus-specific PCR detection for identification of Aeromonas in tissue samples to assist the medico-legal diagnosis of death by drowning

Huys, Geert; Coopman, Vera; Varenbergh, Dirk Van; Cordonnier, Jan

doi:10.1016/j.forsciint.2012.03.017

Cited by 20 publications

(14 citation statements)

References 33 publications

(42 reference statements)

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“…Microorganisms can be detected by incubation in selective media or by various PCR techniques. In addition to DNA fragments specific for some aquatic bacteria, the 16S ribosomal RNA gene (16 rDNA) has been repeatedly used as a target for bacterial and planktonic elements (He et al 2008;Tie et al 2010;Huys et al 2012;Kakizaki et al 2012). …”

Section: Microbiological Testsmentioning

confidence: 99%

Injuries due to Asphyxiation and Drowning

Keil¹,

Lunetta²,

Vann³

et al. 2014

Handbook of Forensic Medicine

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Section: Microbiological Testsmentioning

confidence: 99%

Injuries due to Asphyxiation and Drowning

Keil¹,

Lunetta²,

Vann³

et al. 2014

Handbook of Forensic Medicine

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“…; Huys et al . ), but there was no report about the detection of four genera bacteria at one assay. The proposed mPCR system was designed not to detect specific species, but to detect and discriminate bacteria of four specific target genera.…”

Section: Resultsmentioning

confidence: 99%

“…A sensitive, specific and costeffective detection technique is strongly needed to monitor fish populations and detect carrier fish. The application of PCR or mPCR methods for the detection of fish pathogenic bacteria has been previously reported (Del Cerro et al 2002;Mata et al 2004;Picard et al 2004;Panangala et al 2007;Huys et al 2012), but there was no report about the detection of four genera bacteria at one assay. The proposed mPCR system was designed not to detect specific species, but to detect and discriminate bacteria of four specific target genera.…”

Section: Multiplex Pcr Application Of Fish Tissue Samplesmentioning

confidence: 99%

Development of a multiplex PCR assay for rapid and simultaneous detection of four genera of fish pathogenic bacteria

Zhang

2014

Lett Appl Microbiol

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Significance and Impact of the Study: The fish pathogenic bacteria of genus Aeromonas, Vibrio, Edwardsiella and Streptococcus frequently cause severe outbreaks of diseases in cultured fish, and the genus-specific multiplex PCR assay developed in this study can detect the bacteria of the four genera when present in the samples either alone or mixed. The mPCR assay is expected to identify the causative agents more efficiently than uniplex PCR or species-specific multiplex PCR for clinical diagnosis, resulting in the earlier implementation of control measures. This mPCR assay provides a rapid, specific and sensitive tool for the detection or identification of common fish pathogenic bacteria in aquaculture practice. AbstractSpecies of genus Aeromonas, Vibrio, Edwardsiella and Streptococcus are the most common fish pathogenic bacteria that cause economically devastating losses in aquaculture. A multiplex polymerase chain reaction (mPCR) was developed for the simultaneous detection and differentiation of the four genera of fish pathogenic bacteria. Through the use of genus-specific primers instead of species-specific ones, the current mPCR covered much more target bacterial species compared with previously reported species-specific mPCR methods. The specificity of the four putative genus-specific primers was validated experimentally while used exclusively (uniplex PCR) or combined (mPCR) against bacterial genomic DNA templates of the target bacteria and nontarget bacteria. The PCR amplicons for the following genera were obtained as expected: Aeromonas (875 bp), Vibrio (524 bp), Edwardsiella (302 bp) and Streptococcus (197 bp), and the fragments could be separated clearly on the agarose gel electrophoresis. The mPCR did not produce nonspecific amplification products when used to amplify 21 nontarget species of bacteria. The mPCR detection limits for each target bacterial genera were 50 colonyforming units (CFU) in pure culture and 100 CFU in fish tissue samples. In conclusion, the mPCR assay was proven to be a powerful alternative to the conventional culture-based method, given its rapid, specific, sensitive and reliable detection of target pathogens.

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“…However, in rare cases, when identification through diatoms is impractical, other microbes can aid in providing informative results. In the past, methods employed to identify these organisms have included light and electron microscopy, bacterial culture, and PCR (Lunetta et al, 1998;He et al, 2008;Lucci et al, 2008;Tie et al, 2010;Huys et al, 2012;Sitthiwong et al, 2014; Rácz et al, 2016). A culture study with 10 drowning victims and three postmortem submersions revealed fecal coliform (E. coli) and fecal streptococci (E. faecalis) to be useful markers for drowning (Lucci et al, 2008).…”

Section: In Drowningmentioning

confidence: 99%

Microbiome in Death and Beyond: Current Vistas and Future Trends

et al. 2021

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Forensic medicine has, for a long time, been relying on biochemical, anthropologic, and histopathologic evidences in solving various investigations. However, depending on the method used, lengthy sample processing time, scanty sample, and less sensitivity and accuracy pervade these procedures. Accordingly, newer arenas such as the thanatomicrobiome have come forward to aid in its quandaries; furthermore, the parallel advances in genomic and proteomic techniques have complemented and are still emerging to be used in forensic experiments and investigations. Postmortem interval (PMI) is one of the most important aspects of medico-legal investigations. The current trend in PMI estimation is toward genomic analyses of autopsy samples. Similarly, determination of cause of death, although a domain of medical sciences, is being targeted as the next level of forensic casework. With the current trend in laboratory sciences moving to the discovery of newer disease-specific markers for diagnostic and prognostic purposes, the same is being explored for the determination of the cause of death by using techniques such as Real-Time PCR, DNA micro-array, to Next-Gen Sequencing. Establishing an individual’s biological profile has been done using medicolegal methods and anthropology as well as bar-bodies/Davidson bodies (gender determination); and in cases where the determination of age/gender is a challenge using morphological characteristics; the recent advances in the field of genomics and proteomics have played a significant role, e.g., use of mitochondrial DNA in age estimation and in maternity disputes. The major hurdle forensic medical research faces is the fact that most of the studies are conducted in animal models, which are often difficult to mimic in human and real-time scenarios. Additionally, the high accuracy required in criminal investigations to be used in a court of law as evidence has prevented these results to come out of the labs and be used to the optimum. The current review aims at giving a comprehensive and critical account of the various molecular biology techniques including “thanatogenomics,” currently being utilized in the veritable fields of forensic medicine.

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Selective culturing and genus-specific PCR detection for identification of Aeromonas in tissue samples to assist the medico-legal diagnosis of death by drowning

Cited by 20 publications

References 33 publications

Injuries due to Asphyxiation and Drowning

Injuries due to Asphyxiation and Drowning

Development of a multiplex PCR assay for rapid and simultaneous detection of four genera of fish pathogenic bacteria

Microbiome in Death and Beyond: Current Vistas and Future Trends

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