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PCR Methods Appl. 3:200-204, 1993
©1993 by Cold Spring Harbor Laboratory Press; ISSN 1088-9051
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Genotyping ocular strains of Chlamydia trachomatis by single-tube nested PCR.

S Pecharatana, M A Pickett, P J Watt, and M E Ward

Department of Molecular Microbiology, University Medical School, Southampton General Hospital, UK.

Abstract

A single-tube nested PCR able to discriminate between ocular infection with Chlamydia trachomatis of serovar A, B, or C is described. The method uses genotype-specific primers labeled with different fluorochromes, "drop-in/drop-out" PCR amplification, and product analysis on an Applied Biosystems 373A DNA Sequencer using GENESCAN 672 software. The system readily detected mixed infection with serotypes A and B within the same eye. This strategy provides a rapid genotyping method applicable to a wide variety of epidemiological studies.



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