Detection and discrimination of multiple strains of Zika virus by reverse transcription-loop-mediated isothermal amplification

Tropical Medicine and Health

Seiji Hori¹  Hirotaka Kanuka²  Hiroka Aonuma²  Itoe Iizuka-Shiota³  Tokio Hoshina⁴  Fumihiro Kato⁵  Shigeru Tajima⁵  Masayuki Saijo⁵ 

[1] Department of Infectious Diseases and Infection Control, The Jikei University School of Medicine, Tokyo, Japan;Department of Tropical Medicine, The Jikei University School of Medicine, Tokyo, Japan;Center for Medical Entomology, The Jikei University School of Medicine, Tokyo, Japan;Department of Tropical Medicine, The Jikei University School of Medicine, Tokyo, Japan;Center for Medical Entomology, The Jikei University School of Medicine, Tokyo, Japan;Present Address: Department of Genetics, University of North Carolina at Chapel Hill, Chapel Hill, USA;Department of Tropical Medicine, The Jikei University School of Medicine, Tokyo, Japan;Department of Infectious Diseases and Infection Control, The Jikei University School of Medicine, Tokyo, Japan;Department of Virology 1, National Institute of Infectious Diseases, Tokyo, Japan;

关键词: Zika fever;    Zika virus;    RNA;    Loop-mediated isothermal amplification;    Patient;   

DOI  :  10.1186/s41182-020-00274-z

来源: Springer

【 摘 要 】

BackgroundMonitoring both invasion of Zika virus disease into free countries and circulation in endemic countries is essential to avoid a global pandemic. However, the difficulty lies in detecting Zika virus due to the large variety of mutations in its genomic sequence. To develop a rapid and simple method with high accuracy, reverse transcription-loop-mediated isothermal amplification (RT-LAMP) was adopted for the detection of Zika virus strains derived from several countries.ResultsCommon primers for RT-LAMP were designed based on the genomic sequences of two standard Zika strains: African lineage, MR-766, and Asian lineage, PRVABC59. RT-LAMP reactions using a screened primer set, targeting the NS3 region, detected both Zika virus strains. The minimum detectable quantity was 3 × 10−2 ng of virus RNA. Measurable lag of reaction times among strains was observed. The RT-LAMP method amplified the target virus sequence from the urine and serum of a patient with a travel history in the Caribbean Islands and also provided a prediction about which lineage of Zika virus strain was present.ConclusionsThe RT-LAMP method using a well-optimized primer set demonstrated high specificity and sensitivity for the detection of Zika virus strains with a variety in genomic RNA sequences. In combination with the simplicity of LAMP reaction in isothermal conditions, the optimized primer set established in this study may facilitate rapid and accurate diagnosis of Zika fever patients with virus strain information.

【 授权许可】

CC BY