About: The Development and Application of the Two Real-Time RT-PCR Assays to Detect the Pathogen of HFMD

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type	Academic Article research paper schema:ScholarlyArticle
isDefinedBy	Covid-on-the-Web dataset
title	The Development and Application of the Two Real-Time RT-PCR Assays to Detect the Pathogen of HFMD
Creator	Zhang, Yan Xu, Wenbo Cui, Aili Lu, Yiyu Mao, Naiying »more»
source	PMC
abstract	Large-scale Hand, Foot, and Mouth Disease (HFMD) outbreaks have frequently occurred in China since 2008, affecting more than one million children and causing several hundred children deaths every year. The pathogens of HFMD are mainly human enteroviruses (HEVs). Among them, human enterovirus 71 (HEV71) and coxsackievirus A16 (CVA16) are the most common pathogens of HFMD. However, other HEVs could also cause HFMD. To rapidly detect HEV71 and CVA16, and ensure detection of all HEVs causing HFMD, two real-time hybridization probe-based RT-PCR assays were developed in this study. One is a multiplex real-time RT-PCR assay, which was developed to detect and differentiate HEV71 specifically from CVA16 directly from clinical specimens within 1–2 h, and the other is a broad-spectrum real-time RT-PCR assay, which targeted almost all HEVs. The experiments confirmed that the two assays have high sensitivity and specificity, and the sensitivity was up to 0.1 TCID(50)/ml for detection of HEVs, HEV71, and CVA16, respectively. A total of 213 clinical specimens were simultaneously detected by three kinds of assays, including the two real-time RT-PCR assays, direct conventional RT-PCR assay, and virus isolation assay on human rhabdomyosarcoma cells (RD cells). The total positive rate of both HEV71 and CVA16 was 69.48% with real-time RT-PCR assay, 47.42% with RT-PCR assay, and 34.58% with virus isolation assay. One HFMD clinical specimen was positive for HEV, but negative for HEV71 or CVA16, which was identified as Echovirus 11 (Echo11) by virus isolation, RT-PCR, and sequencing for the VP1 gene. The two real-time RT-PCR assays had been applied in 31 provincial HFMD labs to detect the pathogens of HFMD, which has contributed to the rapid identification of the pathogens in the early stages of HFMD outbreaks, and helped to clarify the etiologic agents of HFMD in China.
has issue date	2013-04-18(xsd:dateTime)
bibo:doi	10.1371/journal.pone.0061451
bibo:pmid	23637836
has license	cc-by
sha1sum (hex)	d745e9cbecf81be30e9ffe290781acea84fbd9d3
schema:url	https://doi.org/10.1371/journal.pone.0061451
resource representing a document's title	The Development and Application of the Two Real-Time RT-PCR Assays to Detect the Pathogen of HFMD
has PubMed Central identifier	PMC3630163
has PubMed identifier	23637836
schema:publication	PLoS One
resource representing a document's body	covid:d745e9cbecf81be30e9ffe290781acea84fbd9d3#body_text
is schema:about of	named entity 'broad-spectrum' named entity 'real-time' named entity 'clarify' named entity 'Application' named entity 'DEATHS' »more»

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