Use of whole blood over plasma enhances the detection of dengue virus RNA: possible utility in dengue vaccine trials
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BRIEF REPORT
Use of whole blood over plasma enhances the detection of dengue virus RNA: possible utility in dengue vaccine trials K. Alagarasu1 · M. B. Kakade1 · R. V. Bachal1 · M. Bote1 · D. Parashar1 · P. S. Shah1 Received: 29 July 2020 / Accepted: 4 October 2020 © Springer-Verlag GmbH Austria, part of Springer Nature 2020
Abstract In the present study, the utility of viral RNA isolated from whole blood over plasma for detection of dengue virus (DENV) was investigated in 80 samples referred for serotyping by DENV serotype-specific one-step real-time RT-PCR. DENV RNA was detected in 71.25% of the whole blood samples compared to 46.25% in the corresponding plasma samples. In secondary infections, DENV RNA was detected in 83.3% of whole blood samples, while it was detected in 40.5% of plasma samples (P = 0.0001). Non-structural protein 1 (NS1) antigen was detected in only 54.8% of the secondary infections. The detection rate of DENV RNA in whole blood is higher than in plasma. We suggest that one-step real-time RT-PCR using RNA from whole blood combined with an NS1 ELISA should be the choice for dengue diagnosis in dengue vaccine trials. Early diagnosis of dengue, a disease caused by dengue virus (DENV), depends on the detection of viral RNA or detection of nonstructural protein 1 antigen (NS1 Ag) [1]. The sensitivity for detection of viral RNA and NS1 Ag detection decreases with an increase in the post-onset day of illness. In secondary infection with a heterologous serotype of DENV, the detection window for viral RNA and NS1 Ag is reduced drastically. Serum or plasma is the choice of sample type for diagnosis of dengue [2]. Although IgM capture ELISAs have been reported to be the most useful serological assays in routine dengue diagnosis, the presence of IgM antibodies in subjects immunized with a dengue vaccine might reflect recent vaccination rather than acute DENV infection [3, 4]. Hence, when a vaccine against dengue is available, the diagnosis of dengue will rely exclusively on the detection of viral RNA or NS1 Ag. There is a need, therefore, to look for alternative sample types that can enhance the detection of viral RNA. In the present study, the utility of whole blood
Handling Editor: William G Dundon. Electronic supplementary material The online version of this article (https://doi.org/10.1007/s00705-020-04892-0) contains supplementary material, which is available to authorized users. * K. Alagarasu [email protected]; [email protected] 1
Dengue and Chikungunya Group, ICMR-National Institute of Virology, Pune, Maharashtra 411001, India
compared to plasma for detection of dengue viral RNA was investigated. Under the apex referral laboratory programme for dengue and chikungunya, Indian Council of Medical ResearchNational Institute of Virology (ICMR-NIV), Pune, India receives serum or plasma samples and occasionally whole blood samples in EDTA tubes for detection of dengue virus RNA and antibodies. During the years 2018–2019, 80 whole blood samples were referred from different clinics and hos
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