Molecular detection of Muscovy duck parvovirus by loop-mediated isothermal amplification assay

• Published in: Poultry science Vol. 89; no. 3; pp. 477 - 483
• Main Authors: Ji, J, Xie, Q.M, Chen, C.Y, Bai, S.W, Zou, L.S, Zuo, K.J, Cao, Y.C, Xue, C.Y, Ma, J.Y, Bi, Y.Z
• Format: Journal Article
• Language: English
• Published: Oxford, UK Poultry Science Association 01.03.2010; Oxford University Press
• Subjects: accuracy; Animals; Cairina moschata; coat proteins; cross reaction; disease detection; disease diagnosis; ducks; Ducks - virology; loop-mediated isothermal amplification; microbial detection; microbial genetics; molecular sequence data; Muscovy duck parvovirus; Nucleic Acid Amplification Techniques - methods; Nucleic Acid Amplification Techniques - veterinary; nucleotide sequences; Parvovirus - classification; Parvovirus - isolation & purification; pathogen identification; polymerase chain reaction; Polymerase Chain Reaction - veterinary; Poultry Diseases - virology; rapid methods; Sensitivity and Specificity; test sensitivity; test specificity; loop-mediated isothermal amplification; Muscovy duck parvovirus; molecular detection
• ISSN: 0032-5791; 1525-3171
• DOI: 10.3382/ps.2009-00527

Muscovy duck parvovirus (MDPV) usually causes high morbidity and mortality in 1- to 3-wk-old Muscovy ducklings due to serious infections, which is an imminent threat to the commercial duck industry in China. The objectives of this study were to develop and evaluate a simple, rapid, and inexpensive loop-mediated isothermal amplification (LAMP) method for specific detection of MDPV and to compare it with the PCR method in rapidity, sensitivity, and accuracy. The novel LAMP assay used a set of 4 specific primers to recognize 6 distinct genomic sequences of capsid protein (VP3) from MDPV, which could be completed within 50 min at 63°C in a simple water bath. The diagnostic results demonstrated that the LAMP assay detected all 7 preserved MDPV isolates, had no cross-reactivity with other duck pathogens (i.e., goose parvovirus, duck plague virus, H9N2 avian influenza virus, duck hepatitis type virus I, and Muscovy duck reovirus). The LAMP assay was at least 10-fold more sensitive than the routine PCR assay and obtained more sensitivity in 61 clinical samples. Therefore, the newly developed LAMP assay provides a specific and sensitive means for detecting MDPV and can be simply applied both in field conditions and in laboratory operations in a cost-effective manner with primary care facilities.