Your browser doesn't support javascript.
loading
Sensitivity and detection of chikungunya viral genetic material using several PCR-based approaches
Esposito, Danillo Lucas Alves; Fonseca, Benedito Antonio Lopes da.
  • Esposito, Danillo Lucas Alves; Universidade de São Paulo. Faculdade de Medicina de Ribeirão Preto. Departamento de Clínica Médica. Ribeirão Preto. BR
  • Fonseca, Benedito Antonio Lopes da; Universidade de São Paulo. Faculdade de Medicina de Ribeirão Preto. Departamento de Clínica Médica. Ribeirão Preto. BR
Rev. Soc. Bras. Med. Trop ; 50(4): 465-469, July-Aug. 2017. tab, graf
Artículo en Inglés | LILACS | ID: biblio-896994
ABSTRACT
Abstract

INTRODUCTION:

Chikungunya fever is a condition resulting from infection by chikungunya virus (CHIKV), an Aedes sp.-transmitted virus. This disease has been diagnosed in thousands of cases in the Americas, particularly in Brazil, in recent years, and there is an ongoing epidemic of chikungunya fever in Brazil that began in 2014. Clinical diagnosis is difficult; only a few cases have been confirmed by laboratory tests due to the low number of specific, efficient tests available for virus or antibody detection. Here, we aimed to evaluate different polymerase chain reaction (PCR) approaches for detection of CHIKV genetic material.

METHODS:

Specific primers and probes within the viral capsid gene region were designed for this work. To evaluate the analytic sensitivity of detection, human sera were spiked with serial dilutions of the viral stock. Several PCR protocols were performed to investigate the sensitivity of CHIKV RNA detection in serum dilutions ranging from 106 to 1 PFU equivalents.

RESULTS:

The technique showing the greatest sensitivity was a real-time PCR assay using specific probes that could detect the genetic material of the virus at all dilutions, followed by conventional PCR. Digital PCR showed low sensitivity and was much more expensive than other technologies. Digital PCR should be used for specific purposes other than clinical diagnosis.

CONCLUSIONS:

Although quantitative PCR using probes was more expensive than the use of intercalating dyes or conventional PCR, it had the highest sensitivity out of all tested PCR approaches.
Asunto(s)


Texto completo: Disponible Índice: LILACS (Américas) Asunto principal: ARN Viral / Virus Chikungunya / Cartilla de ADN / Fiebre Chikungunya Tipo de estudio: Estudio diagnóstico / Guía de Práctica Clínica Límite: Humanos Idioma: Inglés Revista: Rev. Soc. Bras. Med. Trop Asunto de la revista: Medicina Tropical Año: 2017 Tipo del documento: Artículo País de afiliación: Brasil Institución/País de afiliación: Universidade de São Paulo/BR

Similares

MEDLINE

...
LILACS

LIS


Texto completo: Disponible Índice: LILACS (Américas) Asunto principal: ARN Viral / Virus Chikungunya / Cartilla de ADN / Fiebre Chikungunya Tipo de estudio: Estudio diagnóstico / Guía de Práctica Clínica Límite: Humanos Idioma: Inglés Revista: Rev. Soc. Bras. Med. Trop Asunto de la revista: Medicina Tropical Año: 2017 Tipo del documento: Artículo País de afiliación: Brasil Institución/País de afiliación: Universidade de São Paulo/BR