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

Abstract

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 degrees 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.

Isolation and characterization of a distinct duck-origin goose parvovirus causing an outbreak of duckling short beak and dwarfism syndrome in China

“…Death occurs within 2 to 5 days after onset of the disease. MDPV mainly affects young Muscovy ducklings between 2 and 4 weeks old and causes a disease known as 3-W disease, characterized by watery diarrhea, wheezing and locomotor dysfunction, but it does not appear to cause any clinical signs or pathological lesions in goslings or other duckling species [5][6][7][8].…”

Visual detection of goose haemorrhagic polyomavirus in geese and ducks by loop-mediated isothermal amplification

“…Therefore, loop-mediated amplification (LAMP) together with other isothermal techniques including cross-priming amplification or helicase-dependent amplification are exceptionally interesting and useful (Vincent et al 2004;Jeong et al, 2009;Yulong et al, 2010;Xu et al, 2012;Yang et al, 2014;Vincent et al, 2004). The previous report of LAMP application in detection of viral agents of waterfowl showed its robustness in detection of Derzsy's disease virus (GPV; Yang et al, 2010), MDPV (Ji et al, 2010), GCV (Woźniakowski et al, 2012) or DEV (Ji et al, 2009). This report aimed to show its usefulness for GHPV detection.…”

“…An important advance in detection of a number of different human and animal pathogens was the introduction of loop-mediated isothermal amplification (LAMP; Notomi et al, 2000). This method was also recently applied for detection of many avian pathogens (Parida et al, 2004;Huang et al, 2010;Ji et al, 2010;Yang et al, 2010;Woźniakowski et al, 2011). LAMP relies on the isothermal amplification of targeted DNA by Bst, Bsm or GspSSD DNA polymerase with strand-displacement activity.…”

Muscovy Duck Parvovirus Infection with Epicarditis in Bali, Indonesia

“…Clinical and post-mortem findings in MDPVinfected ducklings resemble those of GPV-infected goslings [11], and GPV can cause high mortality in Muscovy ducks as well [16,17]. Further complicating diagnosis of MDPV, the clinical symptoms of MDPV infection are similar to those of not only GPV infection, but also of duck enteritis and duck hepatitis type I infections [18].…”