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Title

Evaluation of an Assay Based On Multiple Detection Temperature Technique for Simultaneous Detection of Viral Gastroenteritis-Causing Pathogens

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Abstract

 Background: Multiple detection temperature (MuDT) technique is an advanced method for the analysis of multiple Ct (cycle threshold) values in a single channel. Objectives: The advantage of this method has been shown only in DNA samples, restricting its diagnostic applicability. This technique was evaluated in this study for its efficacy in the analysis of target RNA. Methods: Allplex GI-virus assay was developed to detect pathogens causing Viral gastroenteritis, one of the major diseases caused by RNAviruses. This one-step multiplex Real-Time Polymerase Chain Reaction (PCR) basedontheMuDTtechnique permits simultaneous amplification and detection of target nucleic acids of norovirus GI, norovirus GII, rotavirus A, adenovirus F, astrovirus, and sapovirus genogroups. The assay was tested for analytical sensitivity, cross-reactivity, repeatability, and applicability to clinical samples. Results: The analytical performance was validated for each target. The assay demonstrated high analytical sensitivity and no crossreactivity, and the repeatability tests showed excellent performance with high accuracy. Analytical performance validation indicated high positive agreement and negative agreement for this method. In the analyses comparing Allplex GI-virus Assay and commercial Seeplex Diarrhea-V ACE Detection using clinical specimens, the positive and negative agreements between the test results were found to be 94. 9% and 98. 8%, respectively. Statistical analysis showed that there was no difference in the performance between the two products. Conclusions: The Allplex GI-virus Assay can rapidly detect six viruses in a single tube without the complementary DNA synthesis step, and this assay was shown to represent an improved molecular diagnostic tool for the simultaneous detection of several RNA viruses. Therefore, our results suggest that theMuDTtechniquemayrepresent anewmolecular diagnostic method for the detection of RNA viruses.

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    APA: Copy

    Han, Jeong Hyun, Kim, Sun Hyung, & Paik, Soon Young. (2017). Evaluation of an Assay Based On Multiple Detection Temperature Technique for Simultaneous Detection of Viral Gastroenteritis-Causing Pathogens. JUNDISHAPUR JOURNAL OF MICROBIOLOGY (JJM), 10(3), 0-0. SID. https://sid.ir/paper/317695/en

    Vancouver: Copy

    Han Jeong Hyun, Kim Sun Hyung, Paik Soon Young. Evaluation of an Assay Based On Multiple Detection Temperature Technique for Simultaneous Detection of Viral Gastroenteritis-Causing Pathogens. JUNDISHAPUR JOURNAL OF MICROBIOLOGY (JJM)[Internet]. 2017;10(3):0-0. Available from: https://sid.ir/paper/317695/en

    IEEE: Copy

    Jeong Hyun Han, Sun Hyung Kim, and Soon Young Paik, “Evaluation of an Assay Based On Multiple Detection Temperature Technique for Simultaneous Detection of Viral Gastroenteritis-Causing Pathogens,” JUNDISHAPUR JOURNAL OF MICROBIOLOGY (JJM), vol. 10, no. 3, pp. 0–0, 2017, [Online]. Available: https://sid.ir/paper/317695/en

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