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Indian Journal of Medical Microbiology
Medknow Publications on behalf of Indian Association of Medical Microbiology
ISSN: 0255-0857 EISSN: 0255-0857
Vol. 23, No. 4, 2005, pp. 239-244
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Bioline Code: mb05072
Full paper language: English
Document type: Research Article
Document available free of charge
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Indian Journal of Medical Microbiology, Vol. 23, No. 4, 2005, pp. 239-244
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Application of polymerase chain reaction to differentiate herpes simplex virus 1 and 2 serotypes in culture negative intraocular aspirates
Shyamal G, Sowmya P, Sudha B, Malathi J, Therese LK, Madhavan HN
Abstract
Purpose: To standardize and apply a polymerase chain reaction (PCR) on the glycoprotein D gene to differentiate Herpes simplex virus (HSV) 1 & 2 serotypes in culture negative intraocular specimens.
Methods: Twenty-one intraocular fluids collected from 19 patients were subjected to cultures for HSV and uniplex PCR (uPCR) for DNA polymerase gene. To differentiate HSV serotypes, as 1 & 2, a seminested PCR (snPCR) targeting the glycoprotein D gene was standardised and applied onto 21 intraocular fluids. The specificity of the snPCR was verified by application onto ATCC strains of HSV 1 and 2, clinical isolates and DNA sequencing of the amplified products. All specimens were also tested for the presence of cytomegalovirus (CMV) and varicella zoster virus (VZV) by nucleic acid amplification methods.
Results: Four of the 21 intraocular fluids were positive for HSV by uPCR. snPCR detected HSV in three additional specimens (total of seven specimens), and identified three as HSV 1 and four as HSV 2. DNA sequencing of PCR products showed 100% homology with the standard strains of HSV 1 and 2 respectively. None of the samples were positive in culture. Among the other patients, CMV DNA was detected in two and VZV DNA in five others.
Conclusions: The standardized snPCR can be applied directly onto the culture negative specimens for rapid differentiation of HSV serotypes.
Keywords
HSV 1 & 2 serotypes, HSV DNA polymerase gene, HSV Glycoprotein D gene, Viral retinitis, seminested PCR
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© Copyright 2005 Indian Journal of Medical Microbiology. Alternative site location: http://www.ijmm.org
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