Author: Greene, Shermalyn R; Moe, Christine L; Jaykus, Lee-Ann; Cronin, Mike; Grosso, Lynell; Aarle, Pierre van
Title: Evaluation of the NucliSens® Basic Kit assay for detection of Norwalk virus RNA in stool specimens Cord-id: 3c1vue0y Document date: 2003_1_16
ID: 3c1vue0y
Snippet: Norwalk-like viruses (NLVs) are a genetically diverse group of human caliciviruses that are the most common cause of epidemic gastroenteritis and are detected typically in stool by reverse transcription (RT)-PCR or electron microscopy (EM). The application of a rapid nucleic acid sequence-based amplification (NASBA) assay for the detection of NLV RNA in stool is described using the NucliSens® Basic Kit. Primers and probes for the NLV Basic Kit assay were based on the RNA polymerase region of th
Document: Norwalk-like viruses (NLVs) are a genetically diverse group of human caliciviruses that are the most common cause of epidemic gastroenteritis and are detected typically in stool by reverse transcription (RT)-PCR or electron microscopy (EM). The application of a rapid nucleic acid sequence-based amplification (NASBA) assay for the detection of NLV RNA in stool is described using the NucliSens® Basic Kit. Primers and probes for the NLV Basic Kit assay were based on the RNA polymerase region of the prototype NLV, Norwalk virus (NV) genome and could consistently detect 10(4) RT-PCR detectable units of NV RNA in a stool filtrate. When compared directly with RT-PCR on a dilution series of NV stool filtrate, the NucliSens® Basic Kit assay was equally sensitive. Cross-reactivity studies with a representative panel of other enteric pathogens were negative. When applied to 15 stool specimens from NV-challenged volunteers, the NASBA Basic Kit application for NV detection yielded 100% sensitivity, 50% specificity, and 67% concordance, using RT-PCR as the ‘gold standard’. Despite the specificity of the NASBA primer/probe sequences for NV, other representatives from both NLV genogroups I and II could be detected by the Basic Kit assay in outbreak stool specimens, although the results were inconsistent. Our results suggest that the NucliSens® Basic Kit assay provides a rapid and sensitive alternative to RT-PCR for detecting NV RNA in stool specimens. However, improvements in test specificity and primer design will be needed before the assay can be used routinely in the clinical setting.
Search related documents:
Co phrase search for related documents- accession number and acid sequence: 1, 2, 3, 4, 5, 6
- accession number and acute phase: 1
- accession number and local alignment blast search tool: 1
- acid amplification and acute phase: 1, 2, 3
- acid amplification and low concentration: 1, 2, 3, 4
- acid sequence and acute phase: 1
- acid sequence and local alignment blast search tool: 1
- acid sequence and low concentration: 1
- acute phase and low concentration: 1, 2, 3
Co phrase search for related documents, hyperlinks ordered by date