A reverse transcription loop-mediated isothermal amplification for broad coverage detection of Asian and African Zika virus lineages

• Published in: BMC infectious diseases Vol. 20; no. 1; pp. 947 - 10
• Main Authors: Teoh, Boon-Teong, Chin, Kim-Ling, Samsudin, Nur-Izyan, Loong, Shih-Keng, Sam, Sing-Sin, Tan, Kim-Kee, Khor, Chee-Sieng, Abd-Jamil, Juraina, Zainal, Nurhafiza, Wilder-Smith, Annelies, Zandi, Keivan, AbuBakar, Sazaly
• Format: Journal Article
• Language: English
• Published: England BioMed Central Ltd 11.12.2020; BioMed Central; BMC
• Subjects: Africa - epidemiology; Antibodies; Asia - epidemiology; Assaying; Cell culture; Cross-reactivity; Diagnostics; Disease control; Disease spread; Epidemics; Evaluation; Gene amplification; Genomes; Guillain-Barre syndrome; Health care; Humans; Infections; Infectious disease; Infectious diseases; Insect control; Laboratories; Marginalized groups; Molecular Diagnostic Techniques - methods; Mosquito; Mosquitoes; Nucleic Acid Amplification Techniques - methods; Nucleic acids; Polymerase chain reaction; Reverse Transcriptase Polymerase Chain Reaction - methods; Reverse transcription; Ribonucleic acid; RNA; RNA, Viral - genetics; RT-LAMP; Sensitivity analysis; Sensitivity and Specificity; Serology; Urine; Vector; Vector-borne; Vector-borne diseases; Viremia; Viruria; Viruses; Zika virus; Zika Virus - classification; Zika Virus - genetics; Zika Virus Infection - diagnosis; Zika Virus Infection - epidemiology; Zika Virus Infection - virology; ZIKV; Asia; Africa; United States > US; Japan; Brazil; Germany; French Polynesia; RT-LAMP; ZIKV; Infectious disease; Diagnostics; Mosquito; Vector; Vector-borne
• ISSN: 1471-2334; 1471-2334
• DOI: 10.1186/s12879-020-05585-4

Early detection of Zika virus (ZIKV) infection during the viremia and viruria facilitates proper patient management and mosquito control measurement to prevent disease spread. Therefore, a cost-effective nucleic acid detection method for the diagnosis of ZIKV infection, especially in resource-deficient settings, is highly required. In the present study, a single-tube reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay was developed for the detection of both the Asian and African-lineage ZIKV. The detection limit, strain coverage and cross-reactivity of the ZIKV RT-LAMP assay was evaluated. The sensitivity and specificity of the RT-LAMP were also evaluated using a total of 24 simulated clinical samples. The ZIKV quantitative reverse transcription-polymerase chain reaction (qRT-PCR) assay was used as the reference assay. The detection limit of the RT-LAMP assay was 3.73 ZIKV RNA copies (probit analysis, P ≤ 0.05). The RT-LAMP assay detected the ZIKV genomes of both the Asian and African lineages without cross-reacting with other arthropod-borne viruses. The sensitivity and specificity of the RT-LAMP assay were 90% (95% CI = 59.6-98.2) and 100% (95% CI = 78.5-100.0), respectively. The RT-LAMP assay detected ZIKV genome in 9 of 24 (37.5%) of the simulated clinical samples compared to 10 of 24 (41.7%) by qRT-PCR assay with a high level of concordance (κ = 0.913, P < 0.001). The RT-LAMP assay is applicable for the broad coverage detection of both the Asian and African ZIKV strains in resource-deficient settings.