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

被引：26

作者：

Vanhomwegen, Jessica ^{[1
]}

Kwasiborski, Aurelia ^{[1
]}

Diop, Abou ^{[2
]}

Boizeau, Laure ^{[3
]}

Hoinard, Damien ^{[1
]}

Vray, Muriel ^{[4
,5
]}

Bercion, Raymond ^{[2
]}

Ndiaye, Babacar ^{[2
]}

Dublineau, Amelie ^{[4
]}

Michiyuki, Satoru ^{[6
]}

Manuguerra, Jean-Claude

Sauvage, Virginie ^{[3
]}

Candotti, Daniel ^{[3
]}

Seck, Abdoulaye ^{[2
,7
]}

Laperche, Syria ^{[3
]}

Shimakawa, Yusuke ^{[4
]}

机构：

[1] Inst Pasteur, Unite Rech & Expertise Environm & Risques Infect, Paris, France

[2] Inst Pasteur, Lab Biol Med, Dakar, Senegal

[3] Inst Natl Transfus Sanguine INTS, Dept Agents Transmissibles Sang, Ctr Natl Reference Risques Infect Transfus, Paris, France

[4] Inst Pasteur, Unit Epidemiol Malad Emergentes, 25-28 Rue Dr Roux, F-75015 Paris, France

[5] Inst Pasteur, Unit Epidemiol Malad Infect, Dakar, Senegal

[6] Eiken Chem Co Ltd, Fundamental Res Lab, Nogi, Tochigi, Japan

[7] Univ Cheikh Anta Diop Dakar, Fac Med Pharm & Odontol, Dakar, Senegal

来源：

CLINICAL MICROBIOLOGY AND INFECTION | 2021年 / 27卷 / 12期

关键词：

Africa; Diagnosis; Hepatitis B; LAMP; Sensitivity and specificity; HEPATITIS-B-VIRUS; INFECTION;

D O I：

10.1016/j.cmi.2021.03.014

中图分类号：

R51 [传染病];

学科分类号：

100401 ;

摘要：

Objective: 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 have developed and evaluated the loop-mediated isothermal amplification (LAMP) assay to diagnose clinically important HBV DNA thresholds defined by the WHO (>20 000 and > 200 000 IU/mL). Methods: 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. Results: 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 and 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. Discussion: We have developed a simple, rapid (60 min), and inexpensive (US$8/assay) alternative to PCR to diagnose high viraemia >= 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. (C) 2021 European Society of Clinical Microbiology and Infectious Diseases. Published by Elsevier Ltd. All rights reserved.

引用

页码：1858.e9 / 1858.e15

页数：7

共 30 条

[1] Detection of Hepatitis B Virus DNA among Chronic and potential Occult HBV patients in resource-limited settings by Loop-Mediated Isothermal Amplification assay [J].