Evaluation of Sensitivity of Multiplex PCR for Detection of
            <i>Mycobacterium tuberculosis</i>
            and
            <i>Pneumocystis jirovecii</i>
            in Clinical Samples

Boondireke, Sirirat; Mungthin, Mathirut; Tan‐ariya, Peerapan; Boonyongsunchai, Petchara; Naaglor, Tawee; Wattanathum, Anan; Treewatchareekorn, Sompong; Leelayoova, Saovanee

doi:10.1128/jcm.00323-10

Cited by 11 publications

(10 citation statements)

References 17 publications

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“…However, reports describing Mrt-PCR approaches for the diagnosis of opportunistic pneumonias are scarce (55). In fact, this is the first report to describe an Mrt-PCR assay for the simultaneous detection of P. jirovecii, H. capsulatum, and C. neoformans.…”

Section: Discussionmentioning

confidence: 98%

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A Multiplex Real-Time PCR Assay for Identification of Pneumocystis jirovecii, Histoplasma capsulatum, and Cryptococcus neoformans/Cryptococcus gattii in Samples from AIDS Patients with Opportunistic Pneumonia

et al. 2014

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A molecular diagnostic technique based on real-time PCR was developed for the simultaneous detection of three of the most frequent causative agents of fungal opportunistic pneumonia in AIDS patients: Pneumocystis jirovecii, Histoplasma capsulatum, and Cryptococcus neoformans/Cryptococcus gattii. This technique was tested in cultured strains and in clinical samples from HIV-positive patients. The methodology used involved species-specific molecular beacon probes targeted to the internal transcribed spacer regions of the rDNA. An internal control was also included in each assay. The multiplex real-time PCR assay was tested in 24 clinical strains and 43 clinical samples from AIDS patients with proven fungal infection. The technique developed showed high reproducibility (r 2 of >0.98) and specificity (100%). For H. capsulatum and Cryptococcus spp., the detection limits of the method were 20 and 2 fg of genomic DNA/20 l reaction mixture, respectively, while for P. jirovecii the detection limit was 2.92 log 10 copies/20 l reaction mixture. The sensitivity in vitro was 100% for clinical strains and 90.7% for clinical samples. The assay was positive for 92.5% of the patients. For one of the patients with proven histoplasmosis, P. jirovecii was also detected in a bronchoalveolar lavage sample. No PCR inhibition was detected. This multiplex real-time PCR technique is fast, sensitive, and specific and may have clinical applications.

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Section: Discussionmentioning

confidence: 98%

“…To our knowledge, only one PCR-based protocol has been previously reported for the detection of tuberculosis and PCP in HIVinfected patients (55). rt-PCR approaches for the detection of C. neoformans/C.…”

Section: Discussionmentioning

confidence: 99%

A Multiplex Real-Time PCR Assay for Identification of Pneumocystis jirovecii, Histoplasma capsulatum, and Cryptococcus neoformans/Cryptococcus gattii in Samples from AIDS Patients with Opportunistic Pneumonia

et al. 2014

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“…The methodology and the cases or samples included are different in each study. Given the importance of the diagnosis of M. tuberculosis, we analysed the outcomes of conventional diagnostic methods and new techniques such as PCR [29]. Table 2 shows four studies that compare and analyse the sensitivity of the PCR.…”

Section: Spinal Tuberculosis Microbiological Diagnosis Methodsmentioning

confidence: 99%