Development and Clinical Validation of Multiplex TaqMan® Assays for Rapid Diagnosis of Viral Gastroenteritis

被引:16
作者
Feeney, S. A. [1 ]
Armstrong, V. J. [1 ]
Mitchell, S. J. [1 ]
Crawford, L. [1 ]
McCaughey, C. [1 ]
Coyle, P. V. [1 ]
机构
[1] Royal Victoria Hosp, Reg Virus Lab, Belfast BT12 6BA, Antrim, North Ireland
关键词
viral gastroenteritis; real-time PCR; REVERSE TRANSCRIPTION-PCR; CAUSATIVE AGENTS; NOROVIRUS; ASTROVIRUS; SAPOVIRUS; ADENOVIRUS; VIRUSES; ROTAVIRUS; DIARRHEA; CHILDREN;
D O I
10.1002/jmv.22162
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
There is a need to provide rapid, sensitive, and often high throughput detection of pathogens in diagnostic virology. Viral gastroenteritis is a serious health issue often leading to hospitalization in the young, the immunocompromised and the elderly. The common causes of viral gastroenteritis include rotavirus, norovirus (genogroups I and II), astrovirus, and group F adenoviruses (serotypes 40 and 41). This article describes the work-up of two internally controlled multiplex, probe-based PCR assays and reports on the clinical validation over a 3-year period, March 2007 to February 2010. Multiplex assays were developed using a combination of TaqMan (TM) and minor groove binder (MGB (TM)) hydrolysis probes. The assays were validated using a panel of 137 specimens, previously positive via a nested gel-based assay. The assays had improved sensitivity for adenovirus, rotavirus, and norovirus (97.3% vs. 86.1%, 100% vs. 87.8%, and 95.1% vs. 79.5%, respectively) and also more specific for targets adenovirus, rotavirus, and norovirus (99% vs. 95.2%, 100% vs. 93.6%, and 97.9% vs. 92.3%, respectively). For the specimens tested, both assays had equal sensitivity and specificity for astrovirus (100%). Overall the probe-based assays detected 16 more positive specimens than the nested gel-based assay. Post-introduction to the routine diagnostic service, a total of 9,846 specimens were processed with multiplex 1 and 2 (7,053 pediatric, 2,793 adult) over the 3-year study period. This clinically validated, probe-based multiplex testing algorithm allows highly sensitive and timely diagnosis of the four most prominent causes of viral gastroenteritis. J. Med. Virol. 83: 1650-1656, 2011. (C) 2011 Wiley-Liss, Inc.
引用
收藏
页码:1650 / 1656
页数:7
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