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Journal of clinical microbiology v.55 no.2, 2017년, pp.526 - 534   SCI SCIE
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Field Validation of SYBR Green- and TaqMan-Based Real-Time PCR Using Biopsy and Swab Samples To Diagnose American Tegumentary Leishmaniasis in an Area Where Leishmania (Viannia) braziliensis Is Endemic

Gomes, Ciro Martins (Division of Dermatology, Department of Medical Clinics, Ribeirááão Preto Medical School, University of Sááãão Paulo, Ribeirááããão Preto, Sááãããão Paulo State, Brazil ) ; Cesetti, Mariana Vicente (Division of Dermatology, Department of Medical Clinics, University of Brasááããããília, Brasááããããíília, Federal District, Brazil ) ; de Paula, Natália Aparecida (Laboratory of Dermatology, University Hospital of Ribeirááããããííão Preto Medical School, University of Sááããããííãão Paulo, Ribeirááããããííããão Preto, Sááããããííãããão Paulo State, ) ; Vernal, Sebastiáán ; Gupta, Gaurav ; Sampaio, Raimunda Nonata Ribeiro ; Roselino, Ana Maria ;
  • 초록  

    The precise diagnosis of American tegumentary leishmaniasis (ATL) is an essential task due to the disease's associated morbidity. A noninvasive, extremely sensitive, and highly specific exam is critical, particularly for mucosal leishmaniasis (ML), in which a low parasite quantity is expected. We aimed to compare the diagnostic accuracy of swab and biopsy sample analysis using SYBR Green- and TaqMan-based real-time PCR (qPCR) assays with that of a composite reference standard consisting of the Montenegro skin test, serology, histopathology, smears, culture, and conventional PCR. In total, 55 patients with ATL (ML, 18 patients; cutaneous leishmaniasis [CL], 37 patients) and 36 patients without ATL were studied. qPCR analysis of swabs was more accurate when using SYBR Green (87.88%; 95% confidence interval [CI], 77.86 to 93.73 patients) than when using TaqMan (78.79%; 95% CI, 67.49 to 86.92%) ( P = 0.031). SYBR Green (84.72%; 95% CI, 74.68 to 91.25%) was also more accurate than TaqMan (73.61%; 95% CI, 62.42 to 82.41%) for biopsy samples ( P = 0.008). All qPCR methods were 100% specific. Swabs and biopsy specimens had similar sensitivity when using the same chemistry ( P = 0.125 for SYBR Green and P = 0.625 for TaqMan). Moreover, qPCR achieved better performance than most existing techniques used for the diagnosis of ATL and also detected the Leishmania parasite in a greater proportion of patients than the associated histopathology, smear, culture, and conventional PCR techniques did. Swabs therefore represent a useful diagnostic tool because they not only are noninvasive but also can achieve an accuracy similar to that of biopsy samples. The high accuracy of SYBR Green-based qPCR may also reduce the requirement for associated parasitological tests for ATL diagnosis.


  • 주제어

    leishmaniasis .   mucocutaneous .   diagnosis .   real-time PCR .   kinetoplast DNA .   real-time polymerase chain reaction.  

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