An Improved Polymerase Cross-Linking Spiral Reaction Assay for Rapid Diagnostic of Canine Parvovirus 2 Infection

Frontiers in Veterinary Science
Xin XuYingzuo Bi

Abstract

With increasing complications of canine parvovirus infection cases, disease diagnosis and treatment have become more difficult. In this study, specificity primers for the conserved region of the VP2 gene of canine parvovirus 2 (CPV-2) were synthesized and evaluated. An improved polymerase cross-linking spiral reaction (PCLSR) method for early and rapid diagnosis of CPV-2 was established. The results showed that the amplification reaction was optimal when run at 62°C for 50 min and could be used to detect CPV-2 without any cross-reactions with other pathogens of canine infectious diseases. Reaction results were directly judged by the naked eyes, with the positive amplification tube shown as luminous yellow and the negative tube as bright purple. Compared with the previously reported polymerase spiral reaction (PSR) method for CPV-2 detection, this reaction was performed using improved primer pairs and a better dye identification method (using an indicator comprising phenol red and cresol red). The detection limit of PCLSR was 3.9 × 101 copies using gel electrophoresis or a visible dye. The positive rate of 132 clinical samples was 42.42%, which was identically the same as that of the PSR method and slightly higher than that of t...Continue Reading

References

Apr 14, 2000·Journal of Veterinary Medicine. B, Infectious Diseases and Veterinary Public Health·J Esfandiari, B Klingeborn
Aug 30, 2008·The Journal of General Virology·Karin HoelzerEdward C Holmes
Feb 16, 2010·Veterinary Research·Karin Hoelzer, Colin R Parrish
Mar 13, 2012·Acta Virologica·M ParthibanD S Bargavi
May 18, 2013·Journal of Virological Methods·André Felipe StreckTimo Homeier
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Jul 9, 2016·The Journal of General Virology·Carla Miranda, Gertrude Thompson
Sep 1, 2018·Journal of Clinical Laboratory Analysis·Minh HoangChao-Nan Lin

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Datasets Mentioned

BETA
GU392236

Methods Mentioned

BETA
gel
PCR
electrophoresis

Software Mentioned

MEGA
Clustal W
SPSS Statistics

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