Detection of Canine Distemper Virus in Blood Samples by Reverse Transcription Loop‐Mediated Isothermal Amplification
- 10 November 2005
- journal article
- Published by Wiley in Journal of Veterinary Medicine, Series B
- Vol. 52 (9), 410-413
- https://doi.org/10.1111/j.1439-0450.2005.00886.x
Abstract
Reverse transcription loop‐mediated isothermal amplification (RT‐LAMP) was used to detect canine distemper virus (CDV) genomic RNA. A set of four primers, two outer and two inner, were designed from CDV genomic RNA targeting the nucleocapsid protein gene. The optimal reaction time and temperature for LAMP were determined to be 60 min at 65°C. The relative sensitivity and specificity of RT‐LAMP was found to be 100% and 93.3%, respectively, based on 50 canine blood samples and using RT‐PCR as the gold standard. The detection limit of the RT‐LAMP method was 100 times lower than with RT‐PCR (10‐1TCID50 ml−1 versus 10TCID50 ml−1). In addition to the advantage resulting from the visual detection of the end‐product, the LAMP method is fast, requiring only 1 h to complete the assay. The LAMP method is a viable alternative to RT‐PCR for diagnosing CDV infection in dogs. The LAMP method might be useful as an on site diagnostic assay for detecting CDV.Keywords
This publication has 14 references indexed in Scilit:
- Development and Evaluation of a Novel Loop-Mediated Isothermal Amplification Method for Rapid Detection of Severe Acute Respiratory Syndrome CoronavirusJournal of Clinical Microbiology, 2004
- Comparison of one‐step RT‐PCR and a nested PCR for the detection of canine distemper virus in clinical samplesAustralian Veterinary Journal, 2004
- Apoptosis in the Cerebellum of Dogs with DistemperJournal of Veterinary Medicine, Series B, 2003
- Detection of Japanese yam mosaic virus by RT-LAMPArchiv für die gesamte Virusforschung, 2003
- Detection of Loop-Mediated Isothermal Amplification Reaction by Turbidity Derived from Magnesium Pyrophosphate FormationBiochemical and Biophysical Research Communications, 2001
- Loop-mediated isothermal amplification of DNANucleic Acids Research, 2000
- ParamyxoviridaePublished by Elsevier BV ,1999
- Serological Analysis of Canine Distemper Virus Using an Immunocapture ELISA.The Journal of Veterinary Medical Science, 1996
- Distemper in wild carnivores: An epidemiological, histological and immunocytochemical studyVeterinary Microbiology, 1995
- Quantitation of Canine Distemper Virus and Antibodies by Enzyme-Linked Immunosorbent Assays Using Protein A and Monoclonal Antibody CaptureJournal of Veterinary Diagnostic Investigation, 1989