Molecular Detection of Infectious Laryngotracheitis Virus in Chickens with a Microfluidic Chip

Infectious laryngotracheitis (ILT) is a viral disease of chickens’ respiratory system that imposes considerable financial burdens on the chicken industry. Rapid, simple, and specific detection of this virus is crucial to enable proper control measures. Polymerase chain reaction (PCR)-based molecular...

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Bibliographic Details
Published in:Animals (Basel) 2021-11, Vol.11 (11), p.3203
Main Authors: El-Tholoth, Mohamed, Bai, Huiwen, Mauk, Michael G., Anis, Eman, Bau, Haim H.
Format: Article
Language:English
Subjects:
Acids
Assaying
chicken
Chickens
Diagnosis
Gold
Infections
infectious laryngotracheitis virus
Laboratories
Laryngotracheitis
microfluidic device
Microfluidic devices
Polyethylene glycol
Polymerase chain reaction
Poultry farming
quantitative real-time PCR
real-time LAMP
Respiratory system
Sensitivity
Veterinary colleges
Viral diseases
Viruses
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Summary:Infectious laryngotracheitis (ILT) is a viral disease of chickens’ respiratory system that imposes considerable financial burdens on the chicken industry. Rapid, simple, and specific detection of this virus is crucial to enable proper control measures. Polymerase chain reaction (PCR)-based molecular tests require relatively expensive instruments and skilled personnel, confining their application to centralized laboratories. To enable chicken farms to take timely action and contain the spread of infection, we describe a rapid, simple, semi-quantitative benchtop isothermal amplification (LAMP) assay, and a field-deployable microfluidic device for the diagnosis of ILTV infection in chickens. Our assay performance was compared and favorably agreed with quantitative PCR (qPCR). The sensitivity of our real-time LAMP test is 250 genomic copies/reaction. Clinical performance of our microfluidic device using samples from diseased chickens showed 100% specificity and 100% sensitivity in comparison with benchtop LAMP assay and the gold-standard qPCR. Our method facilitates simple, specific, and rapid molecular ILTV detection in low-resource veterinary diagnostic laboratories and can be used for field molecular diagnosis of suspected ILT cases.
ISSN:2076-2615
2076-2615
DOI:10.3390/ani11113203