Universal Single-Probe RT-PCR Assay for Diagnosis of Dengue Virus Infections

Abstract BACKGROUND Dengue virus (DENV) can be transmitted through blood transfusion. DENV was not screened regularly in Xishuangbanna Blood Center. This study was conducted in Xishuangbanna Blood Center with an attempt to develop DENV screening strategies in one of China’s high-incidence areas.METHODS Blood samples were collected randomly between June 2019 and August 2019. These samples were first screened for dengue IgG and IgM antibodies using enzyme-linked immunosorbent assay (ELISA). All reactive samples and some randomly-chosen non-reactive samples were used to detect DENV RNAs using real time polymerase-chain-reaction (RT-PCR) assay. After RT-PCR assay, these samples were further tested for soluble nonstructural protein 1 (NS1) using colloidal gold method. The demographic data of DENV positive donors were collected.RESULTS A total of 2,254 donor samples were collected and tested for dengue IgG and IgM antibodies by ELISA between June 2019 and August 2019. ELISA testing revealed that 598 donor samples were anti-IgG and/or anti-IgM reactive, with a serological prevalence rate of 26.53%. Among all the donor samples, 26 were RT-PCR positive and/or NS1 positive. Moreover, there were significant differences in the prevalence rate of DENV in terms of occupation (P=0.001), education(P<0.001) and ethnicity (P=0.026). CONCLUSION The prevalence of DENV in Xishuangbanna Blood Center was higher than most other blood centers that have implemented DENV donor screening. Our study provides the first-hand data about the prevalence of DENV and allows development of a screening strategy for clinical use.

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Detection of dengue virus serotypes by single-tube multiplex RT-PCR and multiplex real-time PCR assay

Medical Journal Armed Forces India ◽

10.1016/j.mjafi.2021.09.001 ◽

2021 ◽

Author(s):

Kundan Tandel ◽

Mahadevan Kumar ◽

G.S. Bhalla ◽

S.P.S. Shergill ◽

Vijaya Swarnim ◽

...

Keyword(s):

Dengue Virus ◽

Real Time ◽

Real Time Pcr ◽

Rt Pcr ◽

Pcr Assay ◽

Single Tube ◽

Dengue Virus Serotypes

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Prediction of the Spatial Origin of Puumala Virus Infections Using L Segment Sequences Derived from a Generic Screening PCR

Viruses ◽

10.3390/v11080694 ◽

2019 ◽

Vol 11 (8) ◽

pp. 694 ◽

Cited By ~ 3

Author(s):

Weiss ◽

Klempa ◽

Tenner ◽

Kruger ◽

Hofmann

Keyword(s):

Phylogenetic Signal ◽

High Sensitivity ◽

Puumala Virus ◽

Virus Infections ◽

Rt Pcr ◽

Chain Reaction ◽

Pcr Assay ◽

Polymerase Chain ◽

L Segment ◽

Virus Strains

To screen diagnostic specimens for the presence of hantavirus genomes or to identify new hantaviruses in nature, the pan-hanta L-PCR assay, a broadly reactive nested reverse transcription polymerase chain reaction (RT-PCR) assay targeting the L segment, is highly preferred over other assays because of its universality and high sensitivity. In contrast, the geographic allocation of Puumala virus strains to defined outbreak regions in Germany was previously done based on S segment sequences. We show that the routinely generated partial L segment sequences resulting from the pan-hanta L-PCR assay provide sufficient phylogenetic signal to inform the molecular epidemiology of the Puumala virus. Consequently, an additional S segment analysis seems no longer necessary for the identification of the spatial origin of a virus strain.

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P1444 Evaluation of a real-time RT-PCR assay for the diagnosis of measles virus infections during a measles outbreak in Greece

International Journal of Antimicrobial Agents ◽

10.1016/s0924-8579(07)71283-9 ◽

2007 ◽

Vol 29 ◽

pp. S403

Author(s):

S. Kokotas ◽

M. Giannaki ◽

E. Horefti ◽

D. Sgouras ◽

M. Logothetis ◽

...

Keyword(s):

Real Time ◽

Measles Virus ◽

Virus Infections ◽

Rt Pcr ◽

Pcr Assay ◽

Measles Outbreak

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Detection of concurrent infection with multiple dengue virus serotypes in Thai children by ELISA and nested RT-PCR assay

Archives of Virology ◽

10.1007/s00705-008-0249-9 ◽

2008 ◽

Vol 153 (12) ◽

Cited By ~ 15

Author(s):

P. Chinnawirotpisan ◽

M. P. Mammen ◽

A. Nisalak ◽

B. Thaisomboonsuk ◽

S. Narupiti ◽

...

Keyword(s):

Dengue Virus ◽

Concurrent Infection ◽

Rt Pcr ◽

Pcr Assay ◽

Dengue Virus Serotypes

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Detection of Transovarial Transmission on Aedes sp. in Gombong Kebumen Central Java

ASPIRATOR - Journal of Vector-borne Disease Studies ◽

10.22435/asp.v11i2.1476 ◽

2019 ◽

Vol 11 (2) ◽

pp. 107-112

Author(s):

Siwi Pramatama Mars Wijayanti ◽

Devi Octaviana ◽

Arnika Dwi Asti

Keyword(s):

Dengue Virus ◽

Endemic Area ◽

Transovarial Transmission ◽

Next Generation ◽

Rt Pcr ◽

Pcr Assay ◽

Descriptive Research ◽

Central Java ◽

Dengue Transmission ◽

Study Sites

Abstract. Gombong Subdistrict is one of dengue-endemic area in Kebumen Regency, Central Java. The high number of dengue cases in this area raises the question of whether there has been transovarial transmission occured from the Aedes sp. to their eggs. Transovarial transmission could be dangerous because the next generation of mosquitoes can directly become competent vectors as transmitters of the dengue virus (DENV). The purpose of this study is to detect dengue virus transovarial transmission in Aedes sp. in Gombong Subdistrict. This is a descriptive research in Gombong, Semanding, and Kali Tengah villages, Gombong Sub District. A total of 300 houses, 100 houses from each village were selected in this study. There were 600 Oviposition traps (ovitraps) were installed both inside and outside of houses for 6 days. Ovitraps were calculated by its Ovitrap Index (OI). Detection of transovarial transmission was carried by rearing field mosquitoes to Filial 1 then identified by RT-PCR assay. This study showed that OI in the three villages was higher in outdoor compared to indoor positions. All tested samples were negative DENV, indicated that there were no transovarial transmission occured at the study sites. Transmission in these study areas might still through horizontal mechanism transmission by mosquito bites. Although there is no transovarial transmission, awareness of dengue transmission must be continued by eradicating of mosquito nests such as 3M plus activities on a regular basis

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Clinical evaluation of an in-house-developed real-time RT-PCR assay for serotyping of dengue virus

Archives of Virology ◽

10.1007/s00705-020-04725-0 ◽

2020 ◽

Vol 165 (10) ◽

pp. 2311-2315 ◽

Cited By ~ 1

Author(s):

M. B. Kakade ◽

N. Shrivastava ◽

J. A. Patil ◽

D. Parashar ◽

P. S. Shah ◽

...

Keyword(s):

Dengue Virus ◽

Real Time ◽

Clinical Evaluation ◽

Rt Pcr ◽

Pcr Assay

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Reverse Transcriptase PCR (Rt-PCR) for Detection of Dengue and Chikungunya Virus of Mosquito Aedes aegypti in Sokaraja

BioEksakta : Jurnal Ilmiah Biologi Unsoed ◽

10.20884/1.bioe.2020.2.1.1811 ◽

2020 ◽

Vol 2 (1) ◽

pp. 56

Author(s):

Dyah Retno Annisa ◽

Endang Srimurni Kusmintarsih ◽

Trisnowati Budi Ambarningrum

Keyword(s):

Aedes Aegypti ◽

Dengue Virus ◽

Reverse Transcriptase ◽

Chikungunya Virus ◽

Sampling Method ◽

Molecular Detection ◽

Transmission Risk ◽

Virus Infections ◽

Rt Pcr ◽

Cross Sectional

dengue fever (DF). Meanwhile, chikungunya virus causes Chikungunya fever (CF). These diseases involve three organisms, namely virus, mosquito Aedes sp., and human. The transmission of dengue and chikungunya virus is related to the population of Ae. aegypti. Banyumas regency is one of the regions with many cases of dengue and chikungnya virus infections, particularly in Purwokerto, Sokaraja, and Cilongok sub-district. Up to this time, there is no medicine and vaccine provided to treat these viruses effectively. Thus, detection of virus inside vector will be effectively performed in order to predict the transmission risk of dengue and chikungunya virus. This research aimed to know the molecular detection of dengue and chikungunya virus on adult Ae.aegypti mosquito in Sokaraja Region, Banyumas Regency. Survey was done by a cross-sectional method in Sokaraja sub-district from May 2019 – March 2019. Furthermore, technical sampling that used was purposive sampling method of adult Ae.aegypti using BG-Sentital Trap, followed by molecular detection of dengue virus using Two-step RT-PCR and chikungunya gene virus using RT-PCR. Molecular detection of DENV and CHIKV of mosquitoes which collected from Sokaraja region showed negative result

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