Simultaneous Detection of Five Enteric Viruses Associated with Gastroenteritis by Use of a PCR Assay: a Single Real-Time Multiplex Reaction and Its Clinical Application

Jiang, Yixiang; Lin, Fu‐Cheng; Shi, Xiaolu; Zhang, Hailong; Li, Yinghui; Lin, Yinman; Qiu, Yaqun; Chen, Qingliang; Li, Hui; Zhou, Li; Hu, Qinghua

doi:10.1128/jcm.00245-14

Cited by 30 publications

(20 citation statements)

References 27 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…A combination of two probes for Norovirus GI was designed in order to differentiate the five organisms under four detecting channels. The amplification reaction was consistent with previously described and cycling condition was the same as that of simultaneous detecting virulence loci of DEC or five enteric bacteria (Jiang et al, 2014). …”

Section: Methodssupporting

confidence: 54%

“…Viral RNA was reverse transcribed for 10 min at 30°C, 50 min at 42°C, and 5 min at 95°C. A single real-time multiplex PCR assay was followed for simultaneous detection of five enteric viruses: norovirus genogroups I and II; sapovirus genogroups I, II, IV, and V; human rotavirus A; adenovirus serotypes 40 and 41; and human astrovirus (Jiang et al, 2014). The specific primers and probes used in this reaction were presented in Table 1 .…”

Section: Methodsmentioning

confidence: 99%

See 1 more Smart Citation

The 12 Gastrointestinal Pathogens Spectrum of Acute Infectious Diarrhea in a Sentinel Hospital, Shenzhen, China

Shen

Zhang

et al. 2016

Front. Microbiol.

Self Cite

View full text Add to dashboard Cite

Acute infectious gastroenteritis is one of the most common diseases among all ages, particularly in developing countries. The pathogen spectrum may differ among different regions and seasons. To investigate the etiology of acute diarrhea in Shenzhen, a prospective study was conducted from August 2014 to September 2015. Stools from 412 patients with diarrhea (286 of whom were adults) including the general epidemiological information of the patients were collected. The 19 pathogens were detected by conventional culture method or multiplex PCR assay, which included five viruses (rotavirus, adenovirus, sapovirus, norovirus, and astrovirus), 11 bacterial pathogens (Salmonella, Campylobacter jejuni, Shigella, Listeria monocytogenes, Vibrio parahaemolyticus, Vibrio cholera, Enterohemorrhagic (EHEC), enteropathogenic (EPEC), enteroinvasive (EIEC), enterotoxigenic (ETEC); and enteroaggregative Escherichia coli (EAEC)) and three parasites (Entamoeba histolytica, Giardia lamblia, and Cryptosporidium parvum). A potential pathogen and coinfection was found in 41.5 and 7.0% of cases, respectively. The bacterial infection was the dominant cause of diarrhea (32.3%), and the three most frequently identified organisms were Salmonella (12.1%), ETEC (8.0%), and Campylobacter jejuni (4.9%). Salmonella enteritidis was the leading serotype of Salmonella sp. Norovirus (8.3%) and sapovirus (2.2%) were the most common viral pathogens, followed by adenovirus (1.5%) and rotavirus (1.2%). No EHEC, L. monocytogenes, V. cholera, Shigella, and parasites were found. The single most important causes of diarrhea were Salmonella spp. and Campylobacter jejuni, which points toward the need for testing and surveillance for these pathogens in this region.

show abstract

Section: Methodssupporting

confidence: 54%

Section: Methodsmentioning

confidence: 99%

The 12 Gastrointestinal Pathogens Spectrum of Acute Infectious Diarrhea in a Sentinel Hospital, Shenzhen, China

Shen

Zhang

et al. 2016

Front. Microbiol.

Self Cite

View full text Add to dashboard Cite

show abstract

“…Numerous primers with distinct names are quite similar, especially for RdRp-VP1 junction-targeting primer sets (Table 3). Multiplex RT-PCR or PCR assays, whose products were differentiated by agarose gel electrophoresis (101,159,163,169), realtime RT-PCR or PCR (156,158,162,170), and a microspherebased fluorescent PCR product detection assay (e.g., Luminex technology) (160,166), have been reported for the detection of human sapoviruses together with other gastroenteritis viruses. Although these assays aimed for simultaneous detection of multiple viruses, it is unclear whether these assays can detect all genogroups of human sapoviruses.…”

Section: Nucleic Acid Detection Methodsmentioning

confidence: 99%

“…Numerous primers have been designed for the detection of human sapoviruses (Tables 2 to 4). These primers are designed to amplify the partial RdRp (44,46,(141)(142)(143)(144)(146)(147)(148)(149)(150)(151)(152)(153), RdRp-VP1 junction (86,(154)(155)(156)(157)(158)(159)(160)(161)(162), or partial VP1 (126,152,(163)(164)(165)(166) region (Fig. 2).…”

Section: Nucleic Acid Detection Methodsmentioning

confidence: 99%

Comprehensive Review of Human Sapoviruses

et al. 2015

View full text Add to dashboard Cite

SUMMARY Sapoviruses cause acute gastroenteritis in humans and animals. They belong to the genus Sapovirus within the family Caliciviridae . They infect and cause disease in humans of all ages, in both sporadic cases and outbreaks. The clinical symptoms of sapovirus gastroenteritis are indistinguishable from those caused by noroviruses, so laboratory diagnosis is essential to identify the pathogen. Sapoviruses are highly diverse genetically and antigenically. Currently, reverse transcription-PCR (RT-PCR) assays are widely used for sapovirus detection from clinical specimens due to their high sensitivity and broad reactivity as well as the lack of sensitive assays for antigen detection or cell culture systems for the detection of infectious viruses. Sapoviruses were first discovered in 1976 by electron microscopy in diarrheic samples of humans. To date, sapoviruses have also been detected from several animals: pigs, mink, dogs, sea lions, and bats. In this review, we focus on genomic and antigenic features, molecular typing/classification, detection methods, and clinical and epidemiological profiles of human sapoviruses.

show abstract

“…Similarly, a real-time reverse transcription loopmediated isothermal amplification has been developed for rapid and quantitative detection of HAstV-1 (290). Interestingly, some of the published RT-qPCR methods already come in a multiplex format allowing the simultaneous detection of classic HAstVs and other targets such as noroviruses, rotaviruses, sapoviruses, adenoviruses, and/or bocaviruses (291)(292)(293)(294). One of these methods even includes up to 19 targets, including bacteria, protozoa, and helminths (295).…”

Section: Molecular Assaysmentioning

confidence: 99%

Human Astroviruses

2014

View full text Add to dashboard Cite

SUMMARY Human astroviruses (HAtVs) are positive-sense single-stranded RNA viruses that were discovered in 1975. Astroviruses infecting other species, particularly mammalian and avian, were identified and classified into the genera Mamastrovirus and Avastrovirus . Through next-generation sequencing, many new astroviruses infecting different species, including humans, have been described, and the Astroviridae family shows a high diversity and zoonotic potential. Three divergent groups of HAstVs are recognized: the classic ( MAstV 1 ), HAstV-MLB ( MAstV 6 ), and HAstV-VA/HMO ( MAstV 8 and MAstV 9 ) groups. Classic HAstVs contain 8 serotypes and account for 2 to 9% of all acute nonbacterial gastroenteritis in children worldwide. Infections are usually self-limiting but can also spread systemically and cause severe infections in immunocompromised patients. The other groups have also been identified in children with gastroenteritis, but extraintestinal pathologies have been suggested for them as well. Classic HAstVs may be grown in cells, allowing the study of their cell cycle, which is similar to that of caliciviruses. The continuous emergence of new astroviruses with a potential zoonotic transmission highlights the need to gain insights on their biology in order to prevent future health threats. This review focuses on the basic virology, pathogenesis, host response, epidemiology, diagnostic assays, and prevention strategies for HAstVs.

show abstract

Simultaneous Detection of Five Enteric Viruses Associated with Gastroenteritis by Use of a PCR Assay: a Single Real-Time Multiplex Reaction and Its Clinical Application

Cited by 30 publications

References 27 publications

The 12 Gastrointestinal Pathogens Spectrum of Acute Infectious Diarrhea in a Sentinel Hospital, Shenzhen, China

The 12 Gastrointestinal Pathogens Spectrum of Acute Infectious Diarrhea in a Sentinel Hospital, Shenzhen, China

Comprehensive Review of Human Sapoviruses

Human Astroviruses

Contact Info

Product

Resources

About