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Evaluation of a Universal Nested Reverse Transcription Polymerase Chain Reaction for the Detection of Lyssaviruses
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Genetics
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JoVE Journal Genetics
Evaluation of a Universal Nested Reverse Transcription Polymerase Chain Reaction for the Detection of Lyssaviruses

Evaluation of a Universal Nested Reverse Transcription Polymerase Chain Reaction for the Detection of Lyssaviruses

DOI:
10.3791/59428-v

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08:10 min

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May 02, 2019

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, , , , , , ,
1Key Laboratory of Jilin Province for Zoonosis Prevention and Control, Institute of Military Veterinary Medicine,Academy of Military Medical Sciences, 2Jilin Agricultural University, 3Jiangsu Co-innovation Centre for Prevention and Control of Important Animal Infectious Diseases and Zoonoses

Chapters

  • 00:04Title
  • 01:37RNA Extraction and Reverse Transcription of the Viral RNA
  • 03:08First-round PCR
  • 04:03Second-round PCR
  • 04:41Analysis of the PCR Products by Electrophoresis on Agarose Gels
  • 05:48Results: Evaluation of the Reverse Transcription Polymerase Chain Reaction for the Detection of Lyssaviruses
  • 07:44Conclusion

Summary

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A pan-lyssavirus nested reverse transcription polymerase chain reaction has been developed to detect specifically all known lyssaviruses. Validation using rabies brain samples of different animal species showed that this method has a sensitivity and specificity equivalent to the gold standard fluorescent antibody test and could be used for routine rabies diagnosis.

Tags

LyssavirusNested RT-PCRUniversal DetectionAnimal Brain SpecimensBiological Fluid SpecimensRNA ExtractionReverse TranscriptionPCR ContaminationPositive ControlNegative ControlThermocycler

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