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Polymerase chain reaction in the diagnosis of herpetic keratitis: experience in a developing country.
Can J Ophthalmol 2000; 35(3):134-40CJ

Abstract

BACKGROUND

Herpetic ocular disease is a major cause of blindness. Rapid and accurate diagnosis is essential for prompt, proper treatment. We evaluated the usefulness of detection of herpes simplex virus (HSV) DNA by polymerase chain reaction (PCR) in the laboratory diagnosis of herpetic keratitis.

METHODS

A retrospective study was conducted involving 234 patients who attended the cornea clinic at the Regional Ophthalmic Institute, Chennai, India, between March 1995 and September 1997. Inclusion in the study was based on clinical diagnosis of herpetic keratitis. Oligonucleotide primers directed against the HSV-I thymidine kinase gene were used, yielding a 110 base pair amplicon. The utility of PCR analysis was assessed against other diagnostic markers: HSV isolation on cell culture, HSV antigen detection by indirect immunofluorescence, detection of anti-HSV IgG by enzyme-linked immunosorbent assay (ELISA) and detection of HSV-specific tear secretory IgA (sIgA) by ELISA. These tests showed overall sensitivity values of 22.4%, 39.8%, 30.4% and 20.3% respectively.

RESULTS

In epithelial keratitis all 35 specimens from which virus was cultured were positive by PCR. PCR gave a positive result in 23 (82.1%) of the 28 specimens in which HSV antigen was detected and in 4 (57.1%) of the 7 specimens that showed HSV-specific IgG. In addition, PCR detected HSV DNA in 5 of the 30 cases in which these three tests gave a negative result. PCR of two pooled tear samples (collected 1 week apart from the same patient) from 40 patients with stromal keratitis gave a positive result in 12 cases (30%). In stromal keratitis the sensitivity of PCR in detecting HSV DNA in tear samples was 85.7% with culture, indirect immunofluorescence and detection of anti-HSV IgG as the gold standard, and 80% with detection of sIgA as the gold standard.

INTERPRETATION

The results confirm the good correlation with the clinical picture that can be obtained with PCR analysis. They also highlight the diagnostic utility of PCR in detecting HSV DNA in tear samples. This is particularly important in herpetic stromal keratitis, in which collection of corneal scrapings is not advised and, hence, conventional techniques such as virus isolation and antigen detection become difficult.

Authors+Show Affiliations

Department of Microbiology, University of Madras, Taramani, Chennai, India.No affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Comparative Study
Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

10812482

Citation

Pramod, N P., et al. "Polymerase Chain Reaction in the Diagnosis of Herpetic Keratitis: Experience in a Developing Country." Canadian Journal of Ophthalmology. Journal Canadien D'ophtalmologie, vol. 35, no. 3, 2000, pp. 134-40.
Pramod NP, Thyagarajan SP, Mohan KV, et al. Polymerase chain reaction in the diagnosis of herpetic keratitis: experience in a developing country. Can J Ophthalmol. 2000;35(3):134-40.
Pramod, N. P., Thyagarajan, S. P., Mohan, K. V., & Anandakannan, K. (2000). Polymerase chain reaction in the diagnosis of herpetic keratitis: experience in a developing country. Canadian Journal of Ophthalmology. Journal Canadien D'ophtalmologie, 35(3), pp. 134-40.
Pramod NP, et al. Polymerase Chain Reaction in the Diagnosis of Herpetic Keratitis: Experience in a Developing Country. Can J Ophthalmol. 2000;35(3):134-40. PubMed PMID: 10812482.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Polymerase chain reaction in the diagnosis of herpetic keratitis: experience in a developing country. AU - Pramod,N P, AU - Thyagarajan,S P, AU - Mohan,K V, AU - Anandakannan,K, PY - 2000/5/17/pubmed PY - 2000/6/24/medline PY - 2000/5/17/entrez SP - 134 EP - 40 JF - Canadian journal of ophthalmology. Journal canadien d'ophtalmologie JO - Can. J. Ophthalmol. VL - 35 IS - 3 N2 - BACKGROUND: Herpetic ocular disease is a major cause of blindness. Rapid and accurate diagnosis is essential for prompt, proper treatment. We evaluated the usefulness of detection of herpes simplex virus (HSV) DNA by polymerase chain reaction (PCR) in the laboratory diagnosis of herpetic keratitis. METHODS: A retrospective study was conducted involving 234 patients who attended the cornea clinic at the Regional Ophthalmic Institute, Chennai, India, between March 1995 and September 1997. Inclusion in the study was based on clinical diagnosis of herpetic keratitis. Oligonucleotide primers directed against the HSV-I thymidine kinase gene were used, yielding a 110 base pair amplicon. The utility of PCR analysis was assessed against other diagnostic markers: HSV isolation on cell culture, HSV antigen detection by indirect immunofluorescence, detection of anti-HSV IgG by enzyme-linked immunosorbent assay (ELISA) and detection of HSV-specific tear secretory IgA (sIgA) by ELISA. These tests showed overall sensitivity values of 22.4%, 39.8%, 30.4% and 20.3% respectively. RESULTS: In epithelial keratitis all 35 specimens from which virus was cultured were positive by PCR. PCR gave a positive result in 23 (82.1%) of the 28 specimens in which HSV antigen was detected and in 4 (57.1%) of the 7 specimens that showed HSV-specific IgG. In addition, PCR detected HSV DNA in 5 of the 30 cases in which these three tests gave a negative result. PCR of two pooled tear samples (collected 1 week apart from the same patient) from 40 patients with stromal keratitis gave a positive result in 12 cases (30%). In stromal keratitis the sensitivity of PCR in detecting HSV DNA in tear samples was 85.7% with culture, indirect immunofluorescence and detection of anti-HSV IgG as the gold standard, and 80% with detection of sIgA as the gold standard. INTERPRETATION: The results confirm the good correlation with the clinical picture that can be obtained with PCR analysis. They also highlight the diagnostic utility of PCR in detecting HSV DNA in tear samples. This is particularly important in herpetic stromal keratitis, in which collection of corneal scrapings is not advised and, hence, conventional techniques such as virus isolation and antigen detection become difficult. SN - 0008-4182 UR - https://www.unboundmedicine.com/medline/citation/10812482/Polymerase_chain_reaction_in_the_diagnosis_of_herpetic_keratitis:_experience_in_a_developing_country_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0008-4182(00)80006-X DB - PRIME DP - Unbound Medicine ER -