Discrimination of rat-derived Pneumocystis carinii f. sp. carinii and Pneumocystis carinii f. sp. ratti using the polymerase chain reaction

Citation
Rj. Palmer et al., Discrimination of rat-derived Pneumocystis carinii f. sp. carinii and Pneumocystis carinii f. sp. ratti using the polymerase chain reaction, MOL CELL PR, 13(2), 1999, pp. 147-155
Citations number
34
Language
INGLESE
art.tipo
Article
Categorie Soggetti
Molecular Biology & Genetics
Journal title
MOLECULAR AND CELLULAR PROBES
ISSN journal
0890-8508 → ACNP
Volume
13
Issue
2
Year of publication
1999
Pages
147 - 155
Database
ISI
SICI code
0890-8508(199904)13:2<147:DORPCF>2.0.ZU;2-H
Abstract
The rat model of Pneumocystis carinii infection is widely used for the stud y of this non-culturable pathogen. Two genetically divergent 'special forms ' of the organism have been detected in infected rat lungs, P. carinii form ae specialis carinii and P. carinii formae specialis ratti, in some cases a s a co-infection. We have developed a simple and rapid method to analyse ra t-derived P. carinii samples, based on DNA amplification of a portion of th e gene-encoding the mitochondrial large subunit ribosomal RNA. A pair of ol igonucleotide primers were designed for each special form of rat-derived P. carinii, the RC primer pair amplifying a 137 bp fragment from P. carinii f . sp. carinii DNA and the RR primer pair amplifying a 251 bp fragment from P. carinii f. sp. ratti DNA. The specificity of the primers was confirmed b y sequencing the amplification products. The polymerase chain reaction (PCR ) technique was consistent with, and more sensitive than, the electrophoret ic karyotype method. The application of the specific PCR technique has impl ications for future studies on epidemiology, drug sensitivity, immunology a nd molecular biology of rat-derived P. carinii. (C) 1999 Academic Press.