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Development of sensitive and specific loop-mediated isothermal amplification combined with lateral flow device for the rapid detection of hepatitis B virus infection

  • Bacterial, Fungal and Virus Molecular Biology - Research Paper
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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.

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Abbreviations

LAMP:

Loop-mediated isothermal amplification

LFD:

Lateral flow device

LAMP-LFD:

Loop-mediated isothermal amplification-lateral flow device

HBV:

Hepatitis B virus

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Acknowledgements

The authors are thankful to the Genomix Biotech for providing control DNA samples for conducting the research work.

Funding

The complete work was funded and supported by institutional research grants committee.

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Authors and Affiliations

  1. Department of Microbiology, RDGMC, Ujjain, Madhya Pradesh, India

    Soumendra Nath Maity & Yogyata Marothi

  2. Department of Clinical Research Laboratory, Genomix Molecular Diagnostics Pvt. Ltd, Hyderabad, India

    Soumendra Nath Maity, Rudrama Devi Punati, Pratyusha Mallepaddi & Prudhvi Chand Mallepaddi

  3. Department of Medical Microbiology, GenomixCARL Pvt.Ltd, Pulivendula, Andhra Pradesh, India

    Revathi Poonati & Prudhvi Chand Mallepaddi

Authors
  1. Soumendra Nath Maity
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  2. Revathi Poonati
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  3. Rudrama Devi Punati
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  4. Pratyusha Mallepaddi
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  5. Yogyata Marothi
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  6. Prudhvi Chand Mallepaddi
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Contributions

SNM, RP, RDP, and PCM carried out the experimental work and contributed in sample collection. SNM and PM conceptualized the complete work. SNM, RP, and PM contributed in writing and in developing the manuscript. PCM and Yogyata Marothi helped in the design and critically reviewed the manuscript. All the authors read and approved the final copy of the manuscript.

Corresponding author

Correspondence to Soumendra Nath Maity.

Ethics declarations

Ethics approval

The research work was approved by institutional ethical committee of M. N. Raju Medical College and Hospital, Telengana, India conducted on 11-08-2016, approval number ECR/834/Inst/TG/2016.

Conflict of interest

The authors declare no competing interests.

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Responsible Editor: Mauricio Nogueira

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Maity, S.N., Poonati, R., Punati, R.D. et al. Development of sensitive and specific loop-mediated isothermal amplification combined with lateral flow device for the rapid detection of hepatitis B virus infection. Braz J Microbiol 53, 615–623 (2022). https://doi.org/10.1007/s42770-022-00690-9

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  • DOI: https://doi.org/10.1007/s42770-022-00690-9

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