Loop-mediated isothermal amplification: rapid molecular detection of virulence genes associated with avian pathogenic Escherichia coliin poultry

Abstract

Infections with pathogenic Escherichia coli can lead to different animal- and human-associated diseases. E. coli infections are common in intensive poultry farming, and important economic losses can be expected during infections with avian pathogenic E. coli (APEC) strains followed by colibacillosis. Loop-mediated isothermal amplification (LAMP) assays were developed for rapid detection of 3 APEC-associated virulence genes: sitA, traT, and ompT. All 3 LAMP assays are shown to be specific, repeatable, and reproducible. High sensitivities of the assays are shown, where as few as 1,000 bacterial cells/mL can be detected in different matrices. On-site applicability of this LAMP method is demonstrated through testing of different sample types, from animal swabs and tissues, and from environmental samples collected from 6 commercial poultry farms. All 3 virulence genes were detected at high rates (above 85%) in samples from layer and broiler chickens with clinical signs and, interestingly, high prevalence of those genes was detected also in samples collected from clinically healthy broiler flock (above 75%) while lower prevalence was observed in remaining 3 clinically healthy chicken flocks (less than 75%). Importantly, these virulence genes were detected in almost all of the air samples from 11 randomly selected poultry houses, indicating air as an important route of E. coli spread. Three LAMP assays that target APEC-associated virulence genes are shown to be sensitive and robust and are therefore applicable for rapid on-site testing of various sample types, from animal swabs to air. This on-site LAMP testing protocol offers rapid diagnostics, with results obtained in <35 min, and it can be applied to other important microorganisms to allow the required prompt measures to be taken.

Molecular Detection of Avian Pathogenic Escherichia coli (APEC) for the First Time in Layer Farms in Bangladesh and Their Antibiotic Resistance Patterns

“…Previously, Obeng et al [ 58 ] and Stromberg et al [ 49 ] recorded the prevalence of APEC in fecal samples from layers and broilers as 10% and 13%. Kogovšek et al [ 59 ] revealed an association of APEC virulence genes with air samples. This evidence shows that the APEC strains harbored by commercial chickens and their environment can be transmitted from bird to bird and farm to farm through a variety of ways, including air, feces and utensils used in farms.…”

Xiaochaihu Decoction Treatment of Chicken Colibacillosis by Improving Pulmonary Inflammation and Systemic Inflammation

“…Due to the age, resistance, pathogenicity and infection routes of infected chickens, Chicken Colibacillosis include many clinical signs and different disease types. There is a high incidence of diseases in chicken farming, which is responsible for high economic losses in the chicken industry [ 1 , 2 ]. Chicken colibacillosis is clinically characterized by a variety of disease types, including pericarditis, perihepatitis, peritonitis, air sac inflammation and septicemia [ 3 ].…”

Molecular Detection of Some Virulence Genes of E.COLI Isolates From Meat and Meat Product in Port-Said Governorate

“…While Eftekharian et al (2016), detected iutA gene in 70% and lopez banda et al (2014) in 50% of E. coli isolates. The virulence gene (traT) detected in 60% of isolates which nearly to Bok et al (2020) 54.9% while 88.8%and 88.1% detected by Marwa Hassaen et al (2020) and Kogovšek et al (2019). While, 47.7% detected by Ferreira et al (2018).…”