Establishment of multiplex PCR for the detection of common neurotropic viruses
Loading...
Date
2005-03
Authors
Gopalakrishnan, V.
Journal Title
Journal ISSN
Volume Title
Publisher
Pusat Pengajian Sains Perubatan, Universiti Sains Malaysia
Abstract
Viral infections of the central nervous system (CNS) may result in
clinical syndromes like aseptic meningitis, encephalitis and myelitis.
These infections are often difficult to diagnose using conventional
laboratory techniques like viral culture and serology. These
methods are also time consuming and unsatisfactory. Hence a
study was designed to develop a rapid technique to detect the viral
etiology. In this study a reverse transcriptase (RT) multiplex PCR to
detect viral etiologies in CNS infections was standardized. The RT
multiplex PCR was designed to detect enterovirus, herpes simplex
and varicella zoster viruses. Three sets of primers were employed
for their detection. Amplification of target sequences was
qualitatively analyzed by looking for the presence or absence of
amplicons on a 2o/o agarose gel stained with ethidium bromide.
Sensitivity of the PCR has been ascertained. Further, 3 sets of
primers were used to perform a second PCR (nested), which
confirms the product specificity, and also helps in increasing the
sensitivity of the assay. The RT multiplex PCR standardized can be
employed to detect herpes, varicella and enteroviral infections.
Description
Keywords
multiplex PCR