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

Autor: Boon-Teong Teoh, Kim-Ling Chin, Nur-Izyan Samsudin, Shih-Keng Loong, Sing-Sin Sam, Kim-Kee Tan, Chee-Sieng Khor, Juraina Abd-Jamil, Nurhafiza Zainal, Annelies Wilder-Smith, Keivan Zandi, Sazaly AbuBakar
Jazyk: angličtina
Rok vydání: 2020
Předmět:
Zdroj: BMC Infectious Diseases, Vol 20, Iss 1, Pp 1-10 (2020)
Druh dokumentu: article
ISSN: 1471-2334
DOI: 10.1186/s12879-020-05585-4
Popis: Abstract Background 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. Methods 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. Results 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
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