tailieunhanh - Báo cáo hóa học: " Development of a real-time RT-PCR and Reverse Line probe Hybridisation assay for the routine detection and genotyping of Noroviruses in Ireland"

Tham khảo tài liệu 'báo cáo hóa học: " development of a real-time rt-pcr and reverse line probe hybridisation assay for the routine detection and genotyping of noroviruses in ireland"', luận văn - báo cáo phục vụ nhu cầu học tập, nghiên cứu và làm việc hiệu quả | Virology Journal BioMed Central Research Open Access Development of a real-time RT-PCR and Reverse Line probe Hybridisation assay for the routine detection and genotyping of Noroviruses in Ireland John F Menton Karen Kearney and John G Morgan Address Lab 439 Food Science Building Department of Microbiology University College Cork Cork Republic of Ireland Email John F Menton - Karen Kearney - kearney_karen@ John G Morgan - Corresponding author Published 6 September 2007 Received 13 July 2007 Accepted 6 September 2007 Virology Journal 2007 4 86 doi 1743-422X-4-86 This article is available from http content 4 1 86 2007 Menton et al licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License http licenses by which permits unrestricted use distribution and reproduction in any medium provided the original work is properly cited. Abstract Background Noroviruses are the most common cause of non-bacterial gastroenteritis. Improved detection methods have seen a large increase in the number of human NoV genotypes in the last ten years. The objective of this study was to develop a fast method to detect quantify and genotype positive NoV samples from Irish hospitals. Results A real-time RT-PCR assay and a Reverse Line Blot Hybridisation assay were developed based on the ORF1-ORF2 region. The sensitivity and reactivity of the two assays used was validated using a reference stool panel containing 14 NoV genotypes. The assays were then used to investigate two outbreaks of gastroenteritis in two Irish hospitals. 56 samples were screened for NoV using a real-time RT-PCR assay and 26 samples were found to be positive. Genotyping of these positive samples found that all positives belonged to the GII 4 variant of NoV. Conclusion The combination of the Real-time assay and the reverse line blot hybridisation assay provided a