A novel Approach for Detection of Sindbis Viral RNA with QPCR

Nahla Mohamed, Yazan Odeh

Abstract


SINV was approved as a causative agent of pogosta disease. The seroprevalence of SINV antibodies for the Finnish population is around 2%, considering the prevalence varies between different regions of Finland. While the seroprevalence of SINV antibodies in Sweden highest in central parts of the country. The annual incidence rate in endemic regions of affected countries ranges from 2.7/100,000 in Finland and 2.9/100,000 in Sweden to 18/100,000 in Northern Karelia. This is the most widely distributed of all known arboviruses, affecting all age groups. This study describes the design and evaluation of a rapid and robust quantitative PCR assay able to detect a wide range of different SINV. Primers with the potential to detect all SINV were designed from conserved regions of all different strains of sindbis virus sequences, as identified from multiple alignments. By using SYBR-green-based quantitative real-time PCR (QPCR) protocols, this QPCR assay is able to detect 50-100 target molecules ofsynthetic DNA and less than 100 copies of viral RNA of different SINV. SINV RNA was also detected in clinical samples of patients with SINV has been linked toPogosta diseaseinFinland. Ockelbo is a disease in Sweden and Karelian fever in Russia. The real-time RT-PCR assay is specific and sensitive for detection of SINV and can used for screening SINV in wildlife. This current assay provides a powerful tool for research and diagnostic laboratories where different strains of SINV are circulating worldwide and may be useful in surveys with the purpose of finding new SINV in man and other species.


Keywords


Pogosta disease; SINV; QPCR; mosquito species; bats; ticks; humans; viral RNA.

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References


Brown, F., The classification and nomenclature of viruses: summary of results of meetings of the International Committee on Taxonomy of Viruses in Edmonton, Canada 1987. Intervirology, 1989. 30(4): p. 181-6.

Keegstra, K. and D. Burke, Comparison of the carbohydrate of Sinbis virus glycoproteins with the carbohydrate of host glycoproteins. J Supramol Struct, 1977. 7(3-4): p. 371-9.

Schmaljohn, A.L. and D. McClain, Alphaviruses (Togaviridae) and Flaviviruses (Flaviviridae), in Medical Microbiology, S. Baron, Editor. 1996: Galveston (TX).

Zhou, G., G. Liang, and L. Li, [Complete nucleotide sequence of the nonstructural gene of alphavirus YN87448 strain isolated in China and its relationship to other Sindbis viruses]. Zhonghua Shi Yan He Lin Chuang Bing Du Xue Za Zhi, 1999. 13(4): p. 314-20.

Symington, J. and M.J. Schlesinger, Characterization of a Sinbis virus variant with altered host range. Arch Virol, 1978. 58(2): p. 127-36.

Strauss, E.G., C.M. Rice, and J.H. Strauss, Complete nucleotide sequence of the genomic RNA of Sindbis virus. Virology, 1984. 133(1): p. 92-110.

Blackburn, N.K., et al., Isolation of Sindbis virus from bat organs. Cent Afr J Med, 1982. 28(8): p. 201.

Gresikova, M., et al., Identification of a Sindbis virus strain isolated from Hyaloma marginatum ticks in Sicily. Acta Virol, 1978. 22(3): p. 231-2.

Shabman, R.S., et al., Differential induction of type I interferon responses in myeloid dendritic cells by mosquito and mammalian-cell-derived alphaviruses. J Virol, 2007. 81(1): p. 237-47.

Mukhopadhyay, S., et al., Mapping the structure and function of the E1 and E2 glycoproteins in alphaviruses. Structure, 2006. 14(1): p. 63-73.

Kurkela, S., et al., Causative agent of Pogosta disease isolated from blood and skin lesions. Emerg Infect Dis, 2004. 10(5): p. 889-94.

Brummer-Korvenkontio, M., et al., Epidemiology of Sindbis virus infections in Finland 1981-96: possible factors explaining a peculiar disease pattern. Epidemiol Infect, 2002. 129(2): p. 335-45.

Kurkela, S., et al., Clinical and laboratory manifestations of Sindbis virus infection: prospective study, Finland, 2002-2003. J Infect Dis, 2005. 191(11): p. 1820-9.

Lundstrom, J.O., et al., Geographical and temporal distribution of Ockelbo disease in Sweden. Epidemiol Infect, 1991. 106(3): p. 567-74.

Horling, J., et al., Detection of Ockelbo virus RNA in skin biopsies by polymerase chain reaction. J Clin Microbiol, 1993. 31(8): p. 2004-9.

Kielian, M., Membrane fusion and the alphavirus life cycle. Adv Virus Res, 1995. 45: p. 113-51.

Buckley, A., et al., Serological evidence of West Nile virus, Usutu virus and Sindbis virus infection of birds in the UK. J Gen Virol, 2003. 84(Pt 10): p. 2807-17.

Jamgaonkar, A.V., et al., Serological evidence for Japanese encephalitis virus and West Nile virus infections in water frequenting and terrestrial wild birds in Kolar District, Karnataka State, India. A retrospective study. Acta Virol, 2003. 47(3): p. 185-8.

Muradrasoli, S., et al., Broadly targeted triplex real-time PCR detection of influenza A, B and C viruses based on the nucleoprotein gene and a novel "MegaBeacon" probe strategy. J Virol Methods, 2010. 163(2): p. 313-22.

Escutenaire, S., et al., SYBR Green real-time reverse transcription-polymerase chain reaction assay for the generic detection of coronaviruses. Arch Virol, 2007. 152(1): p. 41-58.

Mohamed, N., et al., A sensitive and quantitative single-tube real-time reverse transcriptase-PCR for detection of enteroviral RNA. J Clin Virol, 2004. 30(2): p. 150-6.

Fokianos, K., I. Sarrou, and I. Pashalidis, Increased radiation exposure by granite used as natural tiling rock in Cypriot houses. Radiation Measurements, 2007. 42(3): p. 446-448.


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