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© Research
Publication : Clinical microbiology and infection : the official publication of the European Society of Clinical Microbiology and Infectious Diseases

Development and clinical validation of loop-mediated isothermal amplification (LAMP) assay to diagnose high HBV DNA levels in resource-limited settings.

Scientific Fields
Diseases
Organisms
Applications
Technique

Published in Clinical microbiology and infection : the official publication of the European Society of Clinical Microbiology and Infectious Diseases - 07 Apr 2021

Vanhomwegen J, Kwasiborski A, Diop A, Boizeau L, Hoinard D, Vray M, Bercion R, Ndiaye B, Dublineau A, Michiyuki S, Manuguerra JC, Sauvage V, Candotti D, Seck A, Laperche S, Shimakawa Y,

Link to Pubmed [PMID] – 33838304

Link to DOI – S1198-743X(21)00149-X10.1016/j.cmi.2021.03.014

Clin Microbiol Infect 2021 Apr; ():

A massive scale-up of testing and treatment is indicated to globally eliminate hepatitis B virus (HBV) infection. However, access to a polymerase chain reaction (PCR), a key test to quantify HBV DNA levels and determine treatment eligibility, is limited in resource-limited countries. We developed and evaluated loop-mediated isothermal amplification (LAMP) assay to diagnose clinically important HBV DNA thresholds defined by WHO (≥20,000 and ≥200,000 IU/mL).Pan-genotypic primer sets were designed on conserved HBV gene regions. Accuracy of LAMP to identify highly viraemic patients was evaluated in 400 and 550 HBV-infected people in France and Senegal, respectively.Our primers successfully detected eight major HBV genotypes/sub-genotypes (A1/2/3/B/C/D/E/F) with a detection limit ranging between 40-400 IU/mL. In France, the area under the receiver operating characteristic curve (AUROC), sensitivity and specificity of bead-based extraction and real-time turbidimetric LAMP were 0.95 (95% CI: 0.93-0.97), 91.1% and 86.0%, respectively, to diagnose HBV DNA ≥20,000 IU/mL; and 0.98 (0.97-0.99), 98.0% and 94.6% for ≥200,000 IU/mL. The performance did not vary by viral genotypes. In Senegal, using a field-adapted method (reagent-free boil-and-spin extraction and inexpensive end-point fluorescence detection), the AUROC, sensitivity and specificity were 0.95 (0.93-0.97), 98.7% and 91.5%, respectively, to diagnose HBV DNA ≥200,000 IU/mL. The assay was not adapted to discriminate low-level viraemia.We developed a simple, rapid (60 minutes), and inexpensive (US$8/assay) alternative to PCR to diagnose high viremia ≥200,000 IU/mL. HBV-LAMP may contribute to eliminating HBV mother-to-child transmission by identifying high-risk pregnant women eligible for antiviral prophylaxis in resource-limited countries.

https://pubmed.ncbi.nlm.nih.gov/33838304