Multilocus Sequence Typing of Pneumocystis jirovecii from Clinical Samples: How Many and Which Loci Should Be Used?

Maitte, C.; Leterrier, Marion; Pape, Patrice Le; Miégeville, M.; Morio, Florent

doi:10.1128/jcm.01073-13

Cited by 54 publications

(86 citation statements)

References 47 publications

(74 reference statements)

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“…Globally, the epidemiology of this disease has always been difficult to study (21,23,28,29), mainly due to the lack of easy-to-perform methods of culture and because alternative powerful and informative techniques have not been available in all microbiology laboratories (30). However, MLST is now used widely to investigate nosocomial PCP outbreaks and is generally considered the gold standard (6,10,11,18,31). Recently, Maitte et al showed that a simple scheme relying on three loci, SOD, mt26S, and CYB, provides sufficient discrimination to be used reliably for epidemiological investigations of PCP outbreaks (11).…”

Section: Discussionmentioning

confidence: 99%

“…However, MLST is now used widely to investigate nosocomial PCP outbreaks and is generally considered the gold standard (6,10,11,18,31). Recently, Maitte et al showed that a simple scheme relying on three loci, SOD, mt26S, and CYB, provides sufficient discrimination to be used reliably for epidemiological investigations of PCP outbreaks (11). Our MLST analysis based on this approach indeed highlighted substantial nucleotide polymorphism in the P. jirovecii genome and evidenced several P. jirovecii infections involving more than one strain (11,18,31,32).…”

Section: Discussionmentioning

confidence: 99%

“…Nucleotide sequencing was achieved using a BigDye Terminator, version 1.1, kit (Life Technologies SAS, Saint-Aubin, France), the primers indicated above, and an ABI-3130XL genetic analyzer (Life Technologies SAS, Saint-Aubin, France). Nucleotide sequences were analyzed using CodonCode Aligner 5.1.5 software (CodonCode Corporation, Centerville, MA) (11,18). The NCBI GenBank accession numbers for the reference sequences are as follows: AF146753 for SOD, M58605 for mt26S, and AF320344 for CYB.…”

Section: Methodsmentioning

confidence: 99%

“…Microsatellite genotyping may be more appropriate for detection of mixed infections (14), and PCR-RFLP analysis of the mitochondrial large-subunit rRNA gene (mtLSU) alone has appeared to be sufficient for roughly distinguishing between strains (12, 13), but MLST has been shown to be highly reproducible and facilitates comparison of data from different laboratories (11).…”

mentioning

confidence: 99%

“…Thus, alternative molecular tools (9, 10), such as multilocus sequence typing (MLST), microsatellite genotyping, or PCR-restriction fragment length polymorphism (PCR-RFLP) analysis, have been used to trace strain circulation in the context of hospitals (11)(12)(13). Microsatellite genotyping may be more appropriate for detection of mixed infections (14), and PCR-RFLP analysis of the mitochondrial large-subunit rRNA gene (mtLSU) alone has appeared to be sufficient for roughly distinguishing between strains (12, 13), but MLST has been shown to be highly reproducible and facilitates comparison of data from different laboratories (11).…”

mentioning

confidence: 99%

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Epidemiological Outbreaks of Pneumocystis jirovecii Pneumonia Are Not Limited to Kidney Transplant Recipients: Genotyping Confirms Common Source of Transmission in a Liver Transplantation Unit

et al. 2016

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j Over a 5-month period, four liver transplant patients at a single hospital were diagnosed with Pneumocystis jirovecii pneumonia (PCP). This unusually high incidence was investigated using molecular genotyping. Bronchoalveolar lavage fluids (BALF) obtained from the four liver recipients diagnosed with PCP were processed for multilocus sequence typing (MLST) at three loci (SOD, mt26s, and CYB). Twenty-four other BALF samples, which were positive for P. jirovecii and collected from 24 epidemiologically unrelated patients with clinical signs of PCP, were studied in parallel by use of the same method. Pneumocystis jirovecii isolates from the four liver recipients all had the same genotype, which was different from those of the isolates from all the epidemiologically unrelated individuals studied. These findings supported the hypothesis of a common source of contamination or even cross-transmission of a single P. jirovecii clone between the four liver recipients. Hospitalization mapping showed several possible encounters between these four patients, including outpatient consultations on one particular date when they all possibly met. This study demonstrates the value of molecular genotyping of P. jirovecii isolated from clinical samples for epidemiological investigation of PCP outbreaks. It is also the first description of a common source of exposure to a single P. jirovecii clone between liver transplant recipients and highlights the importance of prophylaxis in such a population.

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