Detection and discrimination of multiple strains of Zika virus by reverse transcription-loop-mediated isothermal amplifi
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(2020) 48:87
Tropical Medicine and Health
RESEARCH
Open Access
Detection and discrimination of multiple strains of Zika virus by reverse transcription-loop-mediated isothermal amplification Hiroka Aonuma1,2, Itoe Iizuka-Shiota1,2,3, Tokio Hoshina1,4, Shigeru Tajima5, Fumihiro Kato5, Seiji Hori4, Masayuki Saijo5 and Hirotaka Kanuka1,2*
Abstract Background: Monitoring 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. Results: Common 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. Conclusions: The 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. Keywords: Zika fever, Zika virus, RNA, Loop-mediated isothermal amplification, Patient
Background Zika virus (ZIKV) is an arbovirus belonging to the family Flaviviridae. ZIKV is transmitted by biting of infected Aedes mosquitoes mainly in tropical regions in the same manner as dengue, chikungunya, and yellow fever. Zika fever (also known as Zika virus disease) became a * Correspondence: [email protected] 1 Department of Tropical Medicine, The Jikei University School of Medicine, Tokyo, Japan 2 Center for Medical Entomology, The Jikei University School of Medicine, Tokyo, Japan Full list of author information is available at the end of the article
globally important medical issue with the declaration of a Public Health Emergency of International Concern by the World Health Organization in February 2016. ZIKV was first identified in 1947 in Uganda, attracting little attention till an association between ZIKV and microcephaly was suspected after a large outbreak in Brazil in 2015 [1, 2]. Most cases of ZIKV infection are thought to be asymptomatic, but some with symptomatic cases showing rash, fever, and arthralgia, and rarely with severe symptomatic Guillain-Barré syndrome. A
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