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However, these methods are limited with respect to the number of target pathogens that can be effectively detected out of a multiplex assay. Several studies have investigated the capability of multiplex RT-PCR and real-time PCR assays to diagnose enteric virus infections, and they showed good sensitivity and specificity. Cell culture and neutralization tests, however, are time-consuming and laborious.
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Many clinical samples remain undetectable due to the absence of antibodies against viral surface proteins. These immunological methods are restricted by the number of pathogens they can detect. Accurate identification of human enteric viruses and early diagnosis from clinical specimens is crucial to patient management and control of infection.Ĭlassical detection of enteric virus is mostly based on conventional approaches, including immunohistochemical detection, indirect immunofluorescence assay (IFA), cell culture, and neutralization testing. Most confirmed cases of EV71-associated HFMD have initial symptoms similar to those of other enteric viral infections but, more frequently, they cause severe neurological complications, including aseptic meningitis, brainstem encephalitis, and acute neurogenic pulmonary edema, none of which are characteristic of enteric viral infection. Despite the close genetic relationship between some enteric viruses, enterovirus 71 (EV71) is generally known to be the major cause of neurological complications and even fatalities in the Asia-Pacific region –. Different species of viruses, including enteroviruses, noroviruses, and astroviruses, have been known to be associated with these diseases.
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The manifestations of human enteric viral infections, which are the most common illnesses during early childhood, range from gastroenteritis to life-threatening diseases such as hand, foot, and mouth disease (HFMD).
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Clinical syndromes are seldom specific to a single pathogen, so accurate and prompt identification of multiplex pathogens has proven invaluable not only to further treatment of the illness, but also to the control of infectious outbreaks and the efficient use of antibiotics and antiviral drugs. Simultaneous detection and classification of all potentially infectious pathogens in a given sample, even when the pathogens cause similar signs and symptoms, is essential to providing the true pathogen spectrum. The identification of specific viruses and clinical specimens shows that the PCR-Mass assay performed as well as or better than standard methods with respect to indicating the presence of multiplex pathogens in a single specimen. The results were compared to those of previous analyses using real-time RT-PCR. Clinical performance was evaluated with a total of 101 clinical specimens from patients suspected of having hand, foot and mouth disease (HFMD). Enteric viral isolates and standard viral RNAs were examined to determine the sensitivity and specificity of the PCR-Mass assay. In this study, a novel PCR-Mass assay combining multiplex polymerase chain reaction (PCR) with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) was developed and used for simultaneous detection of eight distinct human enteric viruses. hand, foot and mouth disease) is essential to the prevention of outbreaks and control of infections. Simultaneous detection of enteric viruses that cause similar symptoms (e.g.