Rapid identification of Pneumocystis carinii isolates by random amplification polymorphic DNA fingerprinting.

Important note: Information in this article was accurate in 1994. The state of the art may have changed since the publication date.

Rapid identification of Pneumocystis carinii isolates by random amplification polymorphic DNA fingerprinting.

Abstr Gen Meet Am Soc Microbiol. 1994;94:622 (abstract no. V-27). Unique Identifier : AIDSLINE ASM94/94313122
Chary-Reddy S; Graves DC; Univ. of Okla. Hlth. Sci. Ctr., Oklahoma City.

Abstract: Pneumocystis carinii is an opportunistic pathogen that causes pneumonia in immunocompromised hosts including humans. It is the leading cause of morbidity and mortality in patients with AIDS. To understand more clearly the transmission of this organism, it is important to develop methods that can distinguish P. carinii isolates. We utilized a randomly designed primer P14 (GCA CAC TCT C) and the random amplified polymorphic DNA (RAPD) assay (Williams et al. 1990. Nucleic acids Res. 18: 6531) to amplify genomic DNA segments of P. carinii obtained from rat, mouse, horse, ferret and human. The amplified products were detected by gel electrophoresis and Southern hybridization using a 32P labelled repetitive DNA sequence from rat P. carinii. Genomic DNA from P. carinii isolates of different species gave distinct RAPD profiles, suggesting that these immunocompromised hosts were infected by genetically distinct P. carinii strains. Genomic DNA of P. carinii obtained from two different strains of rats (Sprague-Dawley from Sasco, Inc. and Wistar from Hilltop laboratories) and Sprague-Dawley rats from different vendors produced different band patterns. Furthermore, we were able to demonstrate two different genetic variants of P. carinii in Wistar rats suggesting a coinfection. In preliminary studies using DNA obtained from two different human P. carinii isolates, identical RAPD profiles were observed. Thus, it appears that the RAPD assay using primer P14 is an attractive method for identifying specific strains of P. carinii and is suitable for large scale epidemiological studies.

Keywords: Animal AIDS-Related Opportunistic Infections/*DIAGNOSIS Base Sequence Blotting, Southern/METHODS Comparative Study DNA Primers Ferrets Gene Amplification/*METHODS Horses Human Mice Molecular Sequence Data Pneumocystis carinii/GENETICS/*ISOLATION & PURIF Pneumocystis carinii Infections/*DIAGNOSIS Polymorphism (Genetics) Random Allocation Rats Rats, Sprague-Dawley Rats, Wistar Repetitive Sequences, Nucleic Acid Species Specificity Variation (Genetics) ABSTRACT
941030
M94A0832

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