- Simultaneous Detection of Waterborne Viruses by Multiplex Real-Time PCR
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Lae-Hyung Kang , Se-hwan Oh , Jeong-Woong Park , Yu-Jung Won , Sangryeol Ryu , Soon-Young Paik
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J. Microbiol. 2013;51(5):671-675. Published online September 14, 2013
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DOI: https://doi.org/10.1007/s12275-013-3199-1
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Norovirus, Rotavirus group A, the Hepatitis A virus, and Coxsackievirus are all common causes of gastroenteritis. Conventional diagnoses of these causative agents are based on antigen detection and electron microscopy. To improve the diagnostic potential for viral gastroenteritis, internally controlled multiplex real-time polymerase chain reaction (PCR) methods have been recently developed. In this study, individual real-time PCRs were developed and optimized for specific detections of Norovirus genogroup I, Norovirus genogroup II, Rotavirus group A, the Hepatitis A virus, and Coxsackievirus group B1. Subsequently, individual PCRs were combined with multiplex PCR reactions. In general, multiplex real-time PCR assays showed comparable sensitivities and specificities with individual assays. A retrospective clinical evaluation showed increased pathogen detection in 29% of samples using conventional PCR methods. Prospective clinical evaluations were detected in 123 of the 227 (54%) total samples used in the multiplex realtime PCR analysis. The Norovirus genogroup II was found most frequently (23%), followed by Rotavirus (20%), the Hepatitis A virus (4.5%), Coxsackievirus (3.5%), and Norovirus genogroup I (2.6%). Internally controlled multiplex real-time PCR assays for the simultaneous detection of Rotavirus, Coxsackievirus group B, the Hepatitis A virus, and Norovirus genogroups I and II showed significant improvement in the diagnosis of viral gastroenteritis.
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Citations
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Oluwadara Alegbeleye, Anderson S. Sant’Ana
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Lisa Paruch
International Journal of Environmental Research and Public Health.2022; 19(9): 5128. CrossRef - Development and Evaluation of a SYBR Green-Based, Real-time Polymerase Chain Reaction for Rapid and Specific Detection of Human Coxsackievirus B5
Kyu Bong Cho
Biomedical Science Letters.2020; 26(4): 302. CrossRef - Multiplex qPCR for serodetection and serotyping of hepatitis viruses: A brief review
Mohammad Irshad, Priyanka Gupta, Dhananjay Singh Mankotia, Mohammad Ahmad Ansari
World Journal of Gastroenterology.2016; 22(20): 4824. CrossRef - Quantification and Genotyping of Rotavirus A within Two Wastewater Treatment Processes
Chourouk Ibrahim, Nadia Cherif, Salah Hammami, Pierre Pothier, Abdennaceur Hassen
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Applied and Environmental Microbiology.2014; 80(24): 7505. CrossRef
- ppGpp-Mediated Stationary Phase Induction of the Genes Encoded by Horizontally Acquired Pathogenicity Islands and cob/pdu Locus in Salmonella enterica serovar Typhimurium
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Miryoung Song , Hyun-Ju Kim , Sangryeol Ryu , Hyunjin Yoon , Jiae Yun , Hyon E. Choy
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J. Microbiol. 2010;48(1):89-95. Published online March 11, 2010
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DOI: https://doi.org/10.1007/s12275-009-0179-6
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305
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Salmonella enterica is highly diverse in terms of genome structure, which is at least partly due to the horizontal transfer of genetic elements from various sources. In this study, we examined the expression profiles of such genes in Salmonella Pathogenicity Islands (SPIs) and the cob/pdu locus, horizontally acquired large DNA segments, during growth under standard growth conditions. Transcripts from exponentially growing and early stationary phase Salmonellae were compared using various methods including cDNA microarray analysis. Nearly all genes encoded by SPIs and the cob/pdu locus were induced at the onset of the stationary phase in a stringent molecule ppGpp-dependent but stationary phase σ, σ38-independent manner. Although, it has been suggested that ppGpp acts in concert with DksA, we found the stationary phase induction of those SPI genes was not DksA dependent. It is suggested that ppGpp stimulates the expression of these stress-inducible genes encoded by horizontally acquired DNA, by itself or in concert with DksA.
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Citations
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PLOS ONE.2015; 10(6): e0127523. CrossRef - The Primary Transcriptome of Salmonella enterica Serovar Typhimurium and Its Dependence on ppGpp during Late Stationary Phase
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