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Detection and Genogrouping of Noroviruses from Children’s Stools By Taqman One-step RT-PCR
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医学
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JoVE 杂志 医学
Detection and Genogrouping of Noroviruses from Children’s Stools By Taqman One-step RT-PCR

Detection and Genogrouping of Noroviruses from Children’s Stools By Taqman One-step RT-PCR

DOI:
10.3791/3232-v

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15:16 min

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July 22, 2012

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, , , , , , , , ,
1Laboratorio de Investigación y Desarrollo (LID),Universidad Peruana Cayetano Heredia, 2Bloomberg School of Public Health,Johns Hopkins University, 3Laboratorio de Diagnostico Molecular, Facultad de Medicina,University of Concepcion,Chile, 4University of California San Diego School of Medicine

Chapters

  • 00:05Title
  • 01:37In-house Silica-guanidinium Method
  • 02:02Preparation of Silica Suspension for RNA Extraction
  • 03:24Extraction with Guanidine & Silica
  • 06:08Commercial Extraction of Viral RNA: QIAamp Viral RNA Kit
  • 08:32Detection of NoV in Extracted RNA by One-step Real Time PCR with Specific Taqman Probes
  • 14:11Conclusion

Summary

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A One-Step RT-PCR assay for detection and genogroup identification of Norovirus isolates from children’s stools, that utilizes primers and TaqMan probes specific to the open reading frame 1 (ORF1)-ORF2 junction region, the most conserved region of the Norovirus genome is described. A non-commercial, cost-effective RNA extraction method is detailed.

Tags

NorovirusesGenogroupingTaqman One-step RT-PCRAcute GastroenteritisRNA VirusesGenetic DiversityNew StrainsGenogroupsGIGIIGenotypesSubtypesDiagnosisTreatmentRNA ExtractionQIAmp Viral RNA Commercial KitSilica Gel MembraneBuffersMicrospinDiarrheaPathogenesis

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