Development of a colorimetric loop-mediated isothermal amplification assay for rapid and specific detection of Aves polyomavirus 1 from psittacine birds

Loop-Mediated Isothermal Amplification (LAMP) as a Promising Point-of-Care Diagnostic Strategy in Avian Virus Research

“…The incidences of goose hemorrhagic polyomavirus (GHPyV) infection have caused economic losses in waterfowl production at large scale, with a 32% mortality rate of young goslings reported in Poland [ 56 , 60 ]. Adult parrots infected with budgerigar fledgling disease virus (Aves polyomavirus 1, APyV) are more resistant to the inflammatory diseases and while they appear to be asymptomatic, the virus can be shed via feathers and faeces for up to 6 months [ 61 , 62 ]. The fact that the virus can survive in an outside environment for many months raises concerns for swift yet efficient measures such as early screening, cage sanitization and isolation of the infected birds.…”

“…To verify the possibility of any cross-reactivity, the optimized LAMP method was tested against GHPyV with other pathogens present in ducks and geese to observe the potential of cross-reactivity that might lead to a false positive result. Prevalent infections of budgerigar fledgling disease virus (APyV) among parrots are concerning, and should not be neglected in the pet bird industry as well as the global parrot population, which is why the LAMP method has been recently developed for efficient screening of this virus [ 61 ]. Using hydroxyl naphthol blue (HNB) in the reaction mixture for direct monitoring of the results, the colorimetric LAMP method on budgerigar fledgling disease virus has shown 100% sensitivity and specificity in diagnostic performance [ 61 ].…”

“…Prevalent infections of budgerigar fledgling disease virus (APyV) among parrots are concerning, and should not be neglected in the pet bird industry as well as the global parrot population, which is why the LAMP method has been recently developed for efficient screening of this virus [ 61 ]. Using hydroxyl naphthol blue (HNB) in the reaction mixture for direct monitoring of the results, the colorimetric LAMP method on budgerigar fledgling disease virus has shown 100% sensitivity and specificity in diagnostic performance [ 61 ]. The exclusivity of the developed LAMP method was also assessed using different avian pathogens, including the BFDV that commonly co-infects parrots [ 61 ].…”

“…Using hydroxyl naphthol blue (HNB) in the reaction mixture for direct monitoring of the results, the colorimetric LAMP method on budgerigar fledgling disease virus has shown 100% sensitivity and specificity in diagnostic performance [ 61 ]. The exclusivity of the developed LAMP method was also assessed using different avian pathogens, including the BFDV that commonly co-infects parrots [ 61 ]. The limit of APyV detection was observed at 500 copies/reaction for both LAMP and PCR methods [ 61 ].…”

“…The exclusivity of the developed LAMP method was also assessed using different avian pathogens, including the BFDV that commonly co-infects parrots [ 61 ]. The limit of APyV detection was observed at 500 copies/reaction for both LAMP and PCR methods [ 61 ]. The analytical sensitivity results of LAMP being as great as, if not greater than, those from qPCR suggests that the LAMP method is satisfactory to replace the standard diagnostic as a handy and cheaper alternative, especially for diagnostic application in small veterinary clinics or on-field screening.…”