Rapid and sensitive detection ofAvibacterium paragallinarumin the presence of other bacteria using a 5′Taqnuclease assay: a new tool for diagnosing infectious coryza

Abstract

A 5? Taq nuclease assay specific for Avibacterium paragallinarum was designed and optimized for use in diagnosing infectious coryza. The region chosen for assay design was one of known specificity for Av. paragallinarum. The assay detected Av. paragallinarum reference strains representing the three Page and the eight Kume serovars, and field isolates from diverse geographical locations. No cross-reactions were observed with other Avibacterium species, with other bacteria taxonomically related to Av. paragallinarum nor with bacteria and viruses likely to be present in swabs collected from suspected infectious coryza cases. The detection limit for the assay was 6 to 60 colony-forming units per reaction. Twenty-two out of 53 swabs collected from sick birds reacted in the 5? Taq nuclease assay, whereas Av. paragallinarum was not isolated from any of the swabs. All of the 22 swabs yielded other bacteria in culture. The presence of Av. paragallinarum in the swabs was also demonstrated by sequencing, thereby confirming the ability of the assay to detect Av. paragallinarum in the presence of other bacteria. The ability to quantify bacterial load in the swabs using the 5? Taq nuclease assay was demonstrated.

16S ribosomal RNA sequencing and molecular serotyping of Avibacterium paragallinarum isolated from Indian field conditions

“…To overcome this situation, there is a need to use more sensitive and accurate method for diagnosis and confirmation of A. paragallinarum . At present, many alternative approaches such as species-specific polymerase chain reaction (HPG-2 PCR) [ 9 ], DNA Restriction endonuclease analysis [ 10 ], ribotyping [ 11 ], ERIC-PCR [ 12 ], real-time PCR [ 13 ], and 16S ribosomal RNA (rRNA) sequencing [ 7 , 14 ] are employed and found to be more precise and sensitive identifying tools for A. paragallinarum . The serovar level recognition of A. paragallinarum is widely carried out using HA-HI test [ 6 ].…”

Isolation and molecular identification of Avibacterium paragallinarum in suspected cases of infectious coryza

“…More recently, a sensitive and rapid real-time PCR assay was developed for identification of Av. paragallinarum (8).…”

Development of a lateral flow test for the rapid detection of Avibacterium paragallinarum in chickens suspected of having infectious coryza

“…Av. paragallinarum presence in nasal mucus discharge has been detected using molecular techniques such as the TaqMan Real-Time PCR assay [17]. Consequently, we chose to use nasal mucus as a suitable type of sample for lateral flow test evaluation.…”