Abstract
Quickvue and Anigen lateral flow devices (LFDs) were evaluated for detection of H5N1 highly pathogenic avian influenza (HPAI) infections in Egyptian poultry. Sixty five chickens and two turkeys were sampled in eight flocks where H5N1 HPAI infection was suspected. Swabs (tracheal and cloacal) and feathers were collected from each bird for flockside testing by the two LFDs. The same clinical specimens were transported for laboratory testing by M gene RRT PCR where a positive result by this "gold standard" test for one or both swabs from a given bird indicated infection at the bird level, showing 57 birds (including 15 carcassess) to be truly AI infected. Among these 57, similar bird-level LFD testing of swabs showed 43 and 44 to be AI infected by Quickvue and Anigen LFDs, respectively. Nine birds were AI negative by M gene RRT PCR and both LFDs, and one was M gene RRT PCR negative but positive by both LFDs, suggesting one false positive LFD result. Sensitivities of the LFDs relative to M gene RRT PCR were 77.2% for Anigen and 75.4% for Quickvue tests, with 90.0% specificity for both. By including feathers with swabs for LFD testing, the number of LFD positives among 57 infected birds increased by four to 48 by Anigen and 47 by Quickvue, increasing the sensitivity of the LFDs to 84.2% and 82.5% for Anigen and Quickvue, respectively. Although LFD sensitivity cannot compare to the high sensitivity displayed by validated AI RRT PCRs, they may be utilised for flockside testing of birds infected with HPAI at the peak of viral shedding, when birds are displaying advanced clinical signs or sampled as fresh carcasses. Swabs are classic field specimens collected from outbreaks, but inclusion of feathers from birds infected with H5N1 HPAI increased LFD sensitivity. However, the LFD false positive observation emphasises the importance of returning samples for confirmatory laboratory testing.
Highlights
INTRODUCTION(AI) infections in poultry has increased in recent years, Laboratory methods for AI diagnosis are outlined in the following the emergence and extensive spread European Union (EU) and the World Organisation for of H5N1 highly pathogenic (HP) AI (Alexander, 2007; Animal Health (OIE) Diagnostic Manuals, in which virus isolation (VI) in embryonated fowl eggs continues to serve of 2-3 feather calami were tested flockside by each lateral flow devices (LFDs)
Interest in rapid flockside diagnosis of avian influenza health remains a concern (Capua and Alexander, 2007). (AI) infections in poultry has increased in recent years, Laboratory methods for AI diagnosis are outlined in the following the emergence and extensive spread European Union (EU) and the World Organisation for of H5N1 highly pathogenic (HP) AI (Alexander, 2007; Animal Health (OIE) Diagnostic Manuals, in which virus isolation (VI) in embryonated fowl eggs continues to serve of 2-3 feather calami were tested flockside by each lateral flow devices (LFDs)
LFD evaluations have (Spackman et al, 2002) was used as recommended in the been described using field specimens collected during EU AI Diagnostic Manual (EU, 2006) to provide “gold
Summary
(AI) infections in poultry has increased in recent years, Laboratory methods for AI diagnosis are outlined in the following the emergence and extensive spread European Union (EU) and the World Organisation for of H5N1 highly pathogenic (HP) AI (Alexander, 2007; Animal Health (OIE) Diagnostic Manuals, in which virus isolation (VI) in embryonated fowl eggs continues to serve of 2-3 feather calami were tested flockside by each LFD. RNA extraction, AI RRT-PCRs, virus isolation and the potential rapidity with which HPAI typing outbreaks may spread remains a concern. Several RNA was extracted from 140μl of BHIB as previously commercial lateral flow device (LFD) tests are available described (Slomka et al, 2007). Feathers collected from all swabbed being the normal sampling approach during AI outbreak birds were tested by LFDs and M gene RRT-PCR. An individual bird was considered infected if either or both swabs were positive
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