Author/Authors :
Han Jeong-Hyun نويسنده Department of Microbiology, College of Medicine, The
Catholic University of Korea, Seoul, Republic of
Korea , Kim Sun-Hyung نويسنده Seegene Institute of Life Sciences, Seoul, Republic of
Korea , Paik Soon-Young نويسنده Department of Microbiology, College of Medicine, The
Catholic University of Korea, Seoul, Republic of
Korea
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 RNA viruses. This one-step multiplex real-time
polymerase chain reaction (PCR) based on the MuDT technique 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 cross-reactivity,
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 the MuDT
technique may represent a new molecular diagnostic method for the
detection of RNA viruses.
