Soumendra Nath Maity1,2, Revathi Poonati3, Rudrama Devi Punati4, Pratyusha Mallepaddi4, Yogyata Marothi5, Prudhvi Chand Mallepaddi4,3. 1. Department of Microbiology, RDGMC, Ujjain, Madhya Pradesh, India. nath.soumendra@gmail.com. 2. Department of Clinical Research Laboratory, Genomix Molecular Diagnostics Pvt. Ltd, Hyderabad, India. nath.soumendra@gmail.com. 3. Department of Medical Microbiology, GenomixCARL Pvt.Ltd, Pulivendula, Andhra Pradesh, India. 4. Department of Clinical Research Laboratory, Genomix Molecular Diagnostics Pvt. Ltd, Hyderabad, India. 5. Department of Microbiology, RDGMC, Ujjain, Madhya Pradesh, India.
Abstract
Hepatitis B virus is a highly infectious blood borne microbial pathogen that causes several hepatic complications like liver cirrhosis and hepatocellular carcinoma. Several methods are available for the detection of HBV, but every method has their own merits and demerits, which restrict their use in clinical laboratories. The aim of this present study is the development of rapid, inexpensive, sensitive, and specific loop-mediated isothermal amplification followed by lateral flow device (LFD) for detection of HBV in blood specimens. METHODS: HBV standard plasma panels and donor plasma specimens were used to evaluate the assay. HBV DNA was extracted by using QiAamp DNA Blood Mini Kit. Amplification was carried out at constant temperature 63 °C for 60 min. LAMP end products were analyzed by using ESE LAMP tube scanner, gel electrophoresis, UV-lamp, and lateral flow device. RESULTS: HBV-LAMP-LFD assay revealed sensitivity of 92% (138/150) of HBV positive plasma specimens. Specificity of HBV-LAMP-LFD was calculated 100%. CONCLUSION: Our study concludes that HBV-LAMP-LFD is rapid, easy to use, sensitive, and specific point-of-care diagnostic assay for the detection of hepatitis B virus in blood samples. This assay can be used in resource-limited settings as well as in HBV endemic areas.
Hepatitis B virus is a highly infectious blood borne microbial pathogen that causes several hepatic complications like liver cirrhosis and hepatocellular carcinoma. Several methods are available for the detection of HBV, but every method has their own merits and demerits, which restrict their use in clinical laboratories. The aim of this present study is the development of rapid, inexpensive, sensitive, and specific loop-mediated isothermal amplification followed by lateral flow device (LFD) for detection of HBV in blood specimens. METHODS: HBV standard plasma panels and donor plasma specimens were used to evaluate the assay. HBV DNA was extracted by using QiAamp DNA Blood Mini Kit. Amplification was carried out at constant temperature 63 °C for 60 min. LAMP end products were analyzed by using ESE LAMP tube scanner, gel electrophoresis, UV-lamp, and lateral flow device. RESULTS: HBV-LAMP-LFD assay revealed sensitivity of 92% (138/150) of HBV positive plasma specimens. Specificity of HBV-LAMP-LFD was calculated 100%. CONCLUSION: Our study concludes that HBV-LAMP-LFD is rapid, easy to use, sensitive, and specific point-of-care diagnostic assay for the detection of hepatitis B virus in blood samples. This assay can be used in resource-limited settings as well as in HBV endemic areas.
Authors: Angela M Caliendo; Alexander Valsamakis; James W Bremer; Andrea Ferreira-Gonzalez; Suzanne Granger; Linda Sabatini; Gregory J Tsongalis; Yun F Wayne Wang; Belinda Yen-Lieberman; Steve Young; Nell S Lurain Journal: J Clin Microbiol Date: 2011-06-22 Impact factor: 5.948