Heminested Multiplex Reverse Transcription-PCR for Detection and Differentiation of Norwalk-Like Virus Genogroups 1 and 2 in Fecal Samples

AUTOR(ES)

Yuen, Lilly K. W.

FONTE

American Society for Microbiology

RESUMO

The present study describes a heminested multiplex reverse transcription (RT)-PCR assay which enables simultaneous detection and differentiation of Norwalk-like virus (NLV) genogroups from clinical fecal samples without the need to perform sequencing or hybridization. The assay developed was able to detect concentrations of fewer than 100 viral particles per 5 μl of clarified fecal extract and could differentiate the two genogroups with a specificity of 100%. Although the multiplex RT-PCR assay failed to detect NLV in about 3% of the fecal samples which were NLV positive by electron microscopy (EM), the assay was approximately six times more sensitive than EM for NLV detection.

Documentos Relacionados

• Rapid and Efficient Extraction Method for Reverse Transcription-PCR Detection of Hepatitis A and Norwalk-Like Viruses in Shellfish
• Broadly Reactive and Highly Sensitive Assay for Norwalk-Like Viruses Based on Real-Time Quantitative Reverse Transcription-PCR
• Detection and differentiation of antigenically distinct small round-structured viruses (Norwalk-like viruses) by reverse transcription-PCR and southern hybridization.
• Use of heat release and an internal RNA standard control in reverse transcription-PCR detection of Norwalk virus from stool samples.
• Concentration and Detection of Caliciviruses in Water Samples by Reverse Transcription-PCR