NDLI: A real-time RT-PCR for rapid detection and quantification of mosquito-borne alphaviruses

Content Provider	Springer Nature Link
Author	Romeiro, Marilia Farigli Souza, William Marciel Tolardo, Aline Lavado Vieira, Luiz Carlos Henriques, Dyana Alves Araujo, Jansen Siqueira, Carlos Eduardo Hassegawa Colombo, Tatiana Elias Aqui, Victor Hugo Fonseca, Benedito Antonio Lopes Morais Bronzoni, Roberta Vieira gueira, Maurício Lacerda Durigon, Edison Luiz Figueiredo, Luiz Tadeu Moraes
Copyright Year	2016
Abstract	Mosquito-borne alphaviruses are widely distributed throughout the world, causing important human illnesses. Therefore, the development of methods to enable early diagnosis of infections by alphavirus is essential. We show here the development and evaluation of a quantitative real-time RT-PCR using genus-specific primers to the nsP1 viral gene of all mosquito-borne alphaviruses. The specificity and sensitivity of the assay were tested using seven alphaviruses and RNA transcribed from Venezuelan equine encephalitis virus. The detection limits of real-time RT-PCR ranged from 10 to 76 copies per ml. The melting temperature (TM) values for amplification of the alphavirus genomes were 83.05 °C and 85.28 °C. Interestingly, the assay suggested the possibility the arthritogenic alphaviruses with TM peaks of 84.83 to 85.28 °C and encephalitic alphaviruses of 83.34 °C to 84.68 °C could be discriminated both diseases. Real-time RT-PCR may prove very useful for the screening and preliminary diagnosis in outbreaks and surveillance studies as well as for measuring the viral load in pathogenesis studies.
Starting Page	3171
Ending Page	3177
Page Count	7
File Format	PDF
ISSN	03048608
Journal	Archives of Virology
Volume Number	161
Issue Number	11
e-ISSN	14328798
Language	English
Publisher	Springer Vienna
Publisher Date	2016-08-24
Publisher Place	Vienna
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	Virology Medical Microbiology Infectious Diseases
Content Type	Text
Resource Type	Article
Subject	Virology Medicine

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1	Ministry of Education (GoI), Department of Higher Education	Sanctioning Authority	https://www.education.gov.in/ict-initiatives
2	Indian Institute of Technology Kharagpur	Host Institute of the Project: The host institute of the project is responsible for providing infrastructure support and hosting the project	https://www.iitkgp.ac.in
3	National Digital Library of India Office, Indian Institute of Technology Kharagpur	The administrative and infrastructural headquarters of the project	Dr. B. Sutradhar bsutra@ndl.gov.in
4	Project PI / Joint PI	Principal Investigator and Joint Principal Investigators of the project	Dr. B. Sutradhar bsutra@ndl.gov.in Prof. Saswat Chakrabarti will be added soon
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