Direct Viral RNA Detection of SARS-CoV-2 and DENV in Inactivated Samples by Real-Time RT-qPCR: Implications for Diagnosis in Resource Limited Settings with Flavivirus Co-Circulation

被引:3
|
作者
Mao, Zhan Qiu [1 ]
Fukuta, Mizuki [1 ]
Balingit, Jean Claude [1 ]
Nguyen, Thi Thanh Ngan [2 ]
Nguyen, Co Thach [2 ]
Inoue, Shingo [1 ]
Nguyen, Thi Thu Thuy [2 ]
Nguyen, Le Khanh Hang [2 ]
Minakawa, Noboru [1 ]
Morita, Kouichi [1 ]
Le, Thi Quynh Mai [2 ]
Hasebe, Futoshi [1 ]
Moi, Meng Ling [1 ,3 ]
机构
[1] Nagasaki Univ, Inst Trop Med, Grad Sch Biomed Sci, Nagasaki 8528523, Japan
[2] Natl Inst Hyg & Epidemiol, Dept Virol, Hanoi 10000, Vietnam
[3] Univ Tokyo, Sch Int Hlth, Grad Sch Med, Tokyo 1130033, Japan
来源
PATHOGENS | 2021年 / 10卷 / 12期
关键词
SARS-CoV-2; DENV; virus co-circulation; direct RT-qPCR; virus inactivation; biosafety; CORONAVIRUS DISEASE 2019; DENGUE VIRUS TYPE-1; CLINICAL CHARACTERISTICS; SALIVA; INFECTION; URINE; WUHAN;
D O I
10.3390/pathogens10121558
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
The RT-qPCR method remains the gold standard and first-line diagnostic method for the detection of SARS-CoV-2 and flaviviruses, especially in the early stage of viral infection. Rapid and accurate viral detection is a starting point in the containment of the COVID-19 pandemic and flavivirus outbreaks. However, the shortage of diagnostic reagents and supplies, especially in resource-limited countries that experience co-circulation of SARS-CoV-2 and flaviviruses, are limitations that may result in lesser availability of RT-qPCR-based diagnostic tests. In this study, the utility of RNA-free extraction methods was assessed for the direct detection of SARS-CoV-2 and DENV-2 in heat-inactivated or chemical-inactivated samples. The findings demonstrate that direct real-time RT-qPCR is a feasible option in comparison to conventional real-time RT-qPCR based on viral genome extraction-based methods. The utility of heat-inactivation and direct real-time RT-qPCR for SARS-CoV-2, DENV-2 viral RNA detection was demonstrated by using clinical samples of SARS-CoV-2 and DENV-2 and spiked cell culture samples of SARS-CoV-2 and DENV-2. This study provides a simple alternative workflow for flavivirus and SARS-CoV-2 detection that includes heat inactivation and viral RNA extraction-free protocols, with aims to reduce the risk of exposure during processing of SARS-CoV-2 biological specimens and to overcome the supply-chain bottleneck, particularly in resource limited settings with flavivirus co-circulation.
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页数:14
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