Your browser doesn't support javascript.
loading
Design of degenerate primers for multiplex nested-PCR detection of human lymphotropic herpesviruses.
Southeast Asian J Trop Med Public Health ; 2003 Mar; 34(1): 120-5
Artigo em Inglês | IMSEAR | ID: sea-34296
ABSTRACT
To develop the rapid diagnosis and typing of human lymphotropic herpesviruses by using multiplex nested-PCR, the primary PCR (1(o) PCR) primers were redesigned as degenerate primers based on a highly conserved sequences of each DNA polymerase gene of EBV, CMV, HHV-6, HHV-7 and HHV-8. The forward degenerate primer (HHV/1+) contained 12 different sequences, whereas there were 8 different sequences in the reverse degenerate primer (HHV/ 1-). Optimization of multiplex nested-PCR assay conditions were performed to search for the appropriate amount of degenerate primers, dNTP, Taq DNA polymerase, template of secondary PCR (2(o) PCR) and annealing temperature used in 1(o) PCR reaction. Detection sensitivity was the same as described in previous report (approximately 10-100 genome copies). To ensure a true negative result, PCR detection of hepatitis B virus genome was used as internal control. Our presented results, the designed degenerate primers could be used to detect various types of HHV by multiplex nested-PCR.
Assuntos
Texto completo: DisponíveL Índice: IMSEAR (Sudeste Asiático) Assunto principal: Humanos / DNA Viral / Reação em Cadeia da Polimerase / Primers do DNA / Herpesviridae Tipo de estudo: Estudo diagnóstico Idioma: Inglês Revista: Southeast Asian J Trop Med Public Health Ano de publicação: 2003 Tipo de documento: Artigo

Similares

MEDLINE

...
LILACS

LIS

Texto completo: DisponíveL Índice: IMSEAR (Sudeste Asiático) Assunto principal: Humanos / DNA Viral / Reação em Cadeia da Polimerase / Primers do DNA / Herpesviridae Tipo de estudo: Estudo diagnóstico Idioma: Inglês Revista: Southeast Asian J Trop Med Public Health Ano de publicação: 2003 Tipo de documento: Artigo