Polymerase chain reaction with two molecular targets in mucosal leishmaniasis' diagnosis: a validation study

Bracho, Clemencia Ovalle; Quintana, Luisa Porras de; Arenas, Sandra Muvdi; Parra, Melania Rios

doi:10.1590/s0074-02762007005000061

Cited by 22 publications

(16 citation statements)

References 21 publications

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“…Using filter paper as clinical samples, kDNA-targeted PCR was the most sensitive of all the methods used, as corroborated by others 24. kDNA assays are demonstrated to be more sensitive than PCR of rDNA targets due to higher copy number (10, 000 minicircles per parasite as compared with 40–200 copies of the rDNA genes) 6,24,42. Poor performance of ITS2 observed herein might be attributed to PCR inhibition since nested amplification from rDNA PCR was used.…”

Section: Discussionmentioning

confidence: 57%

Cutaneous Leishmaniasis in Khyber Pakhtunkhwa Province of Pakistan: Clinical Diversity and Species-Level Diagnosis

Khan

Bari

Hashim

et al. 2016

The American Journal of Tropical Medicine and Hygiene

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This study primarily aimed to identify the causative species of cutaneous leishmaniasis (CL) in the Khyber Pakhtunkhwa Province of Pakistan and to distinguish any species-specific variation in clinical manifestation of CL. Diagnostic performance of different techniques for identifying CL was assessed. Isolates of Leishmania spp. were detected by in vitro culture, polymerase chain reaction (PCR) on DNA extracted from dried filter papers and microscopic examination of direct lesion smears from patients visiting three major primary care hospitals in Peshawar. A total of 125 CL patients were evaluated. Many acquired the disease from Peshawar and the neighboring tribal area of Khyber Agency. Military personnel acquired CL while deployed in north and south Waziristan. Leishmania tropica was identified as the predominant infecting organism in this study (89.2%) followed by Leishmania major (6.8%) and, unexpectedly, Leishmania infantum (4.1%). These were the first reported cases of CL caused by L. infantum in Pakistan. PCR diagnosis targeting kinetoplast DNA was the most sensitive diagnostic method, identifying 86.5% of all samples found positive by any other method. Other methods were as follows: ribosomal DNA PCR (78.4%), internal transcribed spacer 2 region PCR (70.3%), culture (67.1%), and microscopy (60.5%). Clinical examination reported 14 atypical forms of CL. Atypical lesions were not significantly associated with the infecting Leishmania species, nor with “dry” or “wet” appearance of lesions. Findings from this study provide a platform for species typing of CL patients in Pakistan, utilizing a combination of in vitro culture and molecular diagnostics. Moreover, the clinical diversity described herein can benefit clinicians in devising differential diagnosis of the disease.

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Section: Discussionmentioning

confidence: 57%

Cutaneous Leishmaniasis in Khyber Pakhtunkhwa Province of Pakistan: Clinical Diversity and Species-Level Diagnosis

Khan

Bari

Hashim

et al. 2016

The American Journal of Tropical Medicine and Hygiene

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“…As such, molecular methods have emerged as the new “gold standard” for the diagnosis of cutaneous leishmaniasis 7 , and are particularly useful for forms of the disease that have low parasite loads, such as mucosal leishmaniasis 30 .…”

Section: Discussionmentioning

confidence: 99%

“…According to Volpini et al . 30 , this technique affords differentiation between L. (V.) braziliensis and L. (L.) amazonensis that may otherwise be difficult to determine using cleavage pattern in samples infected with this species.…”

Section: Methodsmentioning

confidence: 99%

Efficiency of noninvasive sampling methods (swab) together with Polymerase Chain Reaction (PCR) for diagnosing American Tegumentary Leishmaniasis

Boni

Oyafuso

Soler

et al. 2017

Rev. Inst. Med. trop. S. Paulo

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Traditional diagnostic methods used to detect American Tegumentary Leishmaniasis, such as histopathology using biopsy samples, culture techniques, and direct search for parasites, have low sensitivity and require invasive collection procedures. This study evaluates the efficiency of noninvasive sampling methods (swab) along with Polymerase Chain Reaction (PCR) for diagnosing American Tegumentary Leishmaniasis using skin and mucous samples from 25 patients who had tested positive for leishmaniasis. The outcome of the tests performance on swab samples was compatible with PCR results on biopsy samples. The findings have also shown that PCR-kDNA test is more efficient than PCR-HSP70 and qPCR tests (sensitivity of 92.3%, 40.7%, and 41%, respectively). Given the high sensitivity of the tests and the fact that the sampling method using swabs affords greater patient comfort and safety, it could be said that this method is a promising alternative to conventional biopsy-based methods for the molecular diagnosis of leishmaniasis.

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“…Indeed, molecular diagnosis provides a new “gold standard” for the diagnosis of dermal leishmaniasis [18], especially for disease presentations that involve a low parasite burden, such as mucosal leishmaniasis [19]. The findings of this study show that the efficiency of recovery and detection of parasite kDNA from swab samples of mucosal tissues analyzed with a combination of PCR using the LV-B1 primer pair and Southern blot hybridization provides noninvasive access to mucosal tissues and a promising diagnostic alternative to biopsy and histopathological evaluation.…”

Section: Discussionmentioning

confidence: 99%

Detection ofLeishmaniain Unaffected Mucosal Tissues of Patients with Cutaneous Leishmaniasis Caused byLeishmania (Viannia)Species

et al. 2009

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Background Leishmania (Viannia) species are the principal cause of mucosal leishmaniasis. The natural history and pathogenesis of mucosal disease are enigmatic. Parasitological evaluation of mucosal tissues has been constrained by the invasiveness of conventional sampling methods. Methods We evaluated the presence ofLeishmania in the mucosa of 26 patients with cutaneous leishmaniasis and 2 patients with mucocutaneous leishmaniasis. Swab samples of the nasal mucosa, tonsils, and conjunctiva were analyzed using polymerase chain reaction with LV-B1 primers and Southern blot hybridization. Results Two patients with mucocutaneous leishmaniasis and 21 (81%) of 26 patients with cutaneous leishmaniasis had Leishmania kinetoplast minicircle DNA (kDNA) in mucosal tissues. kDNA was amplified from swab samples of nasal mucosa from 14 (58%) of 24 patients, tonsils from 13 (46%) of 28 patients, and conjunctiva from 6 (25%) of 24 patients. kDNA was detected in the mucosa of patients with cutaneous disease caused by Leishmania panamensis, Leishmania guyanensis, and Leishmania braziliensis. Conclusion The asymptomatic presence of parasites in mucosal tissues may be common in patients with Leishmania (Viannia) infection.

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Polymerase chain reaction with two molecular targets in mucosal leishmaniasis' diagnosis: a validation study

Abstract: We validated the polymerase chain reaction (PCR)

Cited by 22 publications

References 21 publications

Cutaneous Leishmaniasis in Khyber Pakhtunkhwa Province of Pakistan: Clinical Diversity and Species-Level Diagnosis

Cutaneous Leishmaniasis in Khyber Pakhtunkhwa Province of Pakistan: Clinical Diversity and Species-Level Diagnosis

Efficiency of noninvasive sampling methods (swab) together with Polymerase Chain Reaction (PCR) for diagnosing American Tegumentary Leishmaniasis

Detection ofLeishmaniain Unaffected Mucosal Tissues of Patients with Cutaneous Leishmaniasis Caused byLeishmania (Viannia)Species

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