The Journal of Medical Microbiology, Vol 48, Issue 10 897-905, Copyright © 1999 by Society for General Microbiology

JOURNAL ARTICLE

Oropharyngeal samples for genotyping and monitoring response to treatment in AIDS patients with Pneumocystis carinii pneumonia

A. G. Tsolaki, R. F. Miller and A. E. Wakefield
Department of Paediatrics, Institute of Molecular Medicine, John Radcliffe Hospital, Oxford.

A nested PCR, amplifying a portion of the gene encoding the mitochondrial large subunit ribosomal RNA (mt LSU rRNA) of Pneumocystis carinii sp. f. hominis was applied to oropharyngeal samples obtained on repeated occasions from 12 HIV-infected patients with P. carinii pneumonia (PCP) to monitor response to anti-P. carinii treatment. Genotyping of P. carinii sp. f. hominis was also performed on paired samples of oropharyngeal and broncho-alveolar lavage samples before the start of treatment, and on oropharyngeal samples during the course of treatment, by analysis of sequence variation at the internal transcribed spacer (ITS) regions of the nuclear rRNA operon. When a simple dilutional method was used, a reduction in the amount of amplification product was observed in samples from all patients during the course of treatment. In eight of the 12 patients, a single ITS sequence type was found in the oropharyngeal samples and also in the paired broncho-alveolar lavage sample. A mixed infection was identified in the samples from three patients. In eight patients, the ITS sequence types identified in the oropharyngeal sample were the same as in the broncho-alveolar lavage sample. Nested PCR amplifying the mt LSU rRNA on oropharyngeal samples provides a non-invasive method of monitoring response to treatment of PCP. ITS sequence typing of P. carinii sp. f. hominis from oropharyngeal samples appears to be a reliable alternative to broncho-alveolar lavage samples and provides a non-invasive tool for further epidemiological studies.

This article has been cited by other articles:

				C. Demanche, F. Wanert, M. Barthelemy, J. Mathieu, I. Durand-Joly, E. Dei-Cas, R. Chermette, and J. Guillot Molecular and serological evidence of Pneumocystis circulation in a social organization of healthy macaques (Macaca fascicularis) Microbiology, September 1, 2005; 151(9): 3117 - 3125. [Abstract] [Full Text] [PDF]

				K. Nyamande, U. G. Lalloo, D. York, M. Naidoo, E. M. Irusen, and R. Chetty Low Sensitivity of a Nested Polymerase Chain Reaction in Oropharyngeal Washings for the Diagnosis of Pneumocystis Pneumonia in HIV-Infected Patients Chest, July 1, 2005; 128(1): 167 - 171. [Abstract] [Full Text] [PDF]

				R Miller and L Huang Pneumocystis jirovecii infection Thorax, September 1, 2004; 59(9): 731 - 733. [Full Text] [PDF]

				A. Totet, H. Duwat, E. Magois, V. Jounieaux, P. Roux, C. Raccurt, and G. Nevez Similar genotypes of Pneumocystis jirovecii in different forms of Pneumocystis infection Microbiology, May 1, 2004; 150(5): 1173 - 1178. [Abstract] [Full Text] [PDF]

				F. J. L. Robberts, L. D. Liebowitz, and L. J. Chalkley Genotyping and Coalescent Phylogenetic Analysis of Pneumocystis jiroveci from South Africa J. Clin. Microbiol., April 1, 2004; 42(4): 1505 - 1510. [Abstract] [Full Text] [PDF]

				A. Totet, J.-C. Pautard, C. Raccurt, P. Roux, and G. Nevez Genotypes at the Internal Transcribed Spacers of the Nuclear rRNA Operon of Pneumocystis jiroveci in Nonimmunosuppressed Infants without Severe Pneumonia J. Clin. Microbiol., March 1, 2003; 41(3): 1173 - 1180. [Abstract] [Full Text] [PDF]

				P N Newton, A E Wakefield, R Goldin, and J Govan Pneumocystis carinii pneumonia with pleurisy, platypnoea and orthodeoxia Thorax, February 1, 2003; 58(2): 185 - 186. [Abstract] [Full Text] [PDF]

				G. Nevez, A. Totet, V. Jounieaux, J.-L. Schmit, E. Dei-Cas, and C. Raccurt Pneumocystis jiroveci Internal Transcribed Spacer Types in Patients Colonized by the Fungus and in Patients with Pneumocystosis from the Same French Geographic Region J. Clin. Microbiol., January 1, 2003; 41(1): 181 - 186. [Abstract] [Full Text] [PDF]

				R. F. Miller, H. E. Ambrose, V. Novelli, and A. E. Wakefield Probable Mother-to-Infant Transmission of Pneumocystis carinii f. sp. hominis Infection J. Clin. Microbiol., April 1, 2002; 40(4): 1555 - 1557. [Abstract] [Full Text] [PDF]

				R. F. Miller, H. E. Ambrose, and A. E. Wakefield Pneumocystis carinii f. sp. hominis DNA in Immunocompetent Health Care Workers in Contact with Patients with P. carinii Pneumonia J. Clin. Microbiol., November 1, 2001; 39(11): 3877 - 3882. [Abstract] [Full Text] [PDF]

				Z. Amit and E. S. Kaneshiro Heterogeneity of Pneumocystis Sterol Profiles of Samples from Different Sites in the Same Pair of Lungs Suggests Coinfection by Distinct Organism Populations J. Clin. Microbiol., March 1, 2001; 39(3): 1137 - 1139. [Abstract] [Full Text]

				R. J. Palmer, O. P. Settnes, J. Lodal, and A. E. Wakefield Population Structure of Rat-Derived Pneumocystis carinii in Danish Wild Rats Appl. Envir. Microbiol., November 1, 2000; 66(11): 4954 - 4961. [Abstract] [Full Text]

				N. Maher, S. Vermund, M. Lasbury, C.-H. Lee, M. Bartlett, and T. R. Unnasch Development and Evaluation of a Molecular Viability Assay for Pneumocystis carinii J. Clin. Microbiol., May 1, 2000; 38(5): 1947 - 1952. [Abstract] [Full Text]