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...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
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MDPI AG
2021-11-01
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| Series: | Animals |
| Subjects: | |
| Online Access: | https://www.mdpi.com/2076-2615/11/11/3203 |
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| author | Mohamed El-Tholoth Huiwen Bai Michael G. Mauk Eman Anis Haim H. Bau |
| author_facet | Mohamed El-Tholoth Huiwen Bai Michael G. Mauk Eman Anis Haim H. Bau |
| author_sort | Mohamed El-Tholoth |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-03-10T05:46:08Z |
| format | Article |
| id | doaj.art-68b3ed9b92c74a0e91d2a2007a165a84 |
| institution | Directory Open Access Journal |
| issn | 2076-2615 |
| language | English |
| last_indexed | 2024-03-10T05:46:08Z |
| publishDate | 2021-11-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Animals |
| spelling | doaj.art-68b3ed9b92c74a0e91d2a2007a165a842023-11-22T22:06:59ZengMDPI AGAnimals2076-26152021-11-011111320310.3390/ani11113203Molecular Detection of Infectious Laryngotracheitis Virus in Chickens with a Microfluidic ChipMohamed El-Tholoth0Huiwen Bai1Michael G. Mauk2Eman Anis3Haim H. Bau4Department of Virology, Faculty of Veterinary Medicine, Mansoura University, Mansoura 35516, EgyptDepartment of Mechanical Engineering and Applied Mechanics, University of Pennsylvania, Philadelphia, PA 19104, USADepartment of Mechanical Engineering and Applied Mechanics, University of Pennsylvania, Philadelphia, PA 19104, USANew Bolton Center, Department of Pathobiology, School of Veterinary Medicine, University of Pennsylvania, Kennett Square, PA 19348, USADepartment of Mechanical Engineering and Applied Mechanics, University of Pennsylvania, Philadelphia, PA 19104, USAInfectious 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.https://www.mdpi.com/2076-2615/11/11/3203chickeninfectious laryngotracheitis virusmicrofluidic devicequantitative real-time PCRreal-time LAMP |
| spellingShingle | Mohamed El-Tholoth Huiwen Bai Michael G. Mauk Eman Anis Haim H. Bau Molecular Detection of Infectious Laryngotracheitis Virus in Chickens with a Microfluidic Chip Animals chicken infectious laryngotracheitis virus microfluidic device quantitative real-time PCR real-time LAMP |
| title | Molecular Detection of Infectious Laryngotracheitis Virus in Chickens with a Microfluidic Chip |
| title_full | Molecular Detection of Infectious Laryngotracheitis Virus in Chickens with a Microfluidic Chip |
| title_fullStr | Molecular Detection of Infectious Laryngotracheitis Virus in Chickens with a Microfluidic Chip |
| title_full_unstemmed | Molecular Detection of Infectious Laryngotracheitis Virus in Chickens with a Microfluidic Chip |
| title_short | Molecular Detection of Infectious Laryngotracheitis Virus in Chickens with a Microfluidic Chip |
| title_sort | molecular detection of infectious laryngotracheitis virus in chickens with a microfluidic chip |
| topic | chicken infectious laryngotracheitis virus microfluidic device quantitative real-time PCR real-time LAMP |
| url | https://www.mdpi.com/2076-2615/11/11/3203 |
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