Diagnostic Performance of Filter Paper Lesion Impression PCR for Secondarily Infected Ulcers and Nonulcerative Lesions Caused by Cutaneous Leishmaniasis

Boggild, Andrea K.; Ramos, Ana Pilar; Valencia, Braulio M.; Veland, Nicolás; Calderon, Flor; Arévalo, Jorge; Low, Donald E.; Llanos-Cuentas, Alejandro

doi:10.1128/jcm.02457-10

Cited by 23 publications

(40 citation statements)

References 21 publications

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“…The patient was enrolled in a diagnostic study reported elsewhere, 15 and as such, she underwent lesion scraping, aspiration, and filter paper impression of all three ulcers. Material from lesion scrapings was smeared on glass slides for Giemsa-stained microscopy and assayed for detection of Leishmania (Viannia) kinetoplast DNA (kDNA) by polymerase chain reaction (PCR) as described ( Figure 1A).…”

Section: Case Reportmentioning

confidence: 99%

“…Material from lesion scrapings was smeared on glass slides for Giemsa-stained microscopy and assayed for detection of Leishmania (Viannia) kinetoplast DNA (kDNA) by polymerase chain reaction (PCR) as described ( Figure 1A). 15 Tissue fluid from lesion aspirates was inoculated into traditional Leishmania NovyMacNeal-Nicolle culture medium as described, 16 and assayed by kDNA PCR. Filter paper lesion impressions (FPLIs) were assayed by kDNA PCR as described.…”

Section: Case Reportmentioning

confidence: 99%

“…Filter paper lesion impressions (FPLIs) were assayed by kDNA PCR as described. 15,16 Identification of Leishmania (Viannia) causative species was performed by using mannose phosphate isomerase (mpi), cysteine proteinase B, and heat shock protein 70 PCR and PCR-restriction fragment length polymorphism (PCR-RFLP) analysis according to the algorithm described elsewhere. 17 Specimens from lesions 1 and 2 on the arms amplified product in the mpi PCR when mpi Leishmania (V.) braziliensis primer Figure 1B).…”

Section: Case Reportmentioning

confidence: 99%

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Simultaneous Infection with Leishmania (Viannia) braziliensis and L. (V.) lainsoni in a Peruvian Patient with Cutaneous Leishmaniasis

Veland

Valencia²,

Alba³

et al. 2013

The American Society of Tropical Medicine and Hygiene

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Section: Case Reportmentioning

confidence: 99%

Section: Case Reportmentioning

confidence: 99%

Section: Case Reportmentioning

confidence: 99%

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Simultaneous Infection with Leishmania (Viannia) braziliensis and L. (V.) lainsoni in a Peruvian Patient with Cutaneous Leishmaniasis

Veland

Valencia²,

Alba³

et al. 2013

The American Society of Tropical Medicine and Hygiene

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“…International reference strains and cultured parasites obtained from 80 patients attending our reference center were harvested, and their DNA was extracted as previously described (13). DNAs from anonymized, FPLI specimens from 70 CL lesions collected from 57 Peruvian patients were obtained as reported previously (2,3). Codes and geographical origins are presented in Table S1 in the supplemental material.…”

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confidence: 99%

Leishmania (Viannia) Species Identification on Clinical Samples from Cutaneous Leishmaniasis Patients in Peru: Assessment of a Molecular Stepwise Approach

et al. 2012

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We present an algorithm based on three PCR assays for Leishmania (Viannia) species identification and assessed its performance using 70 specimens from Peruvian patients. The succession of the assayed targets can be ordered according to species prevalence. Sequential progression through the algorithm reduced the number of samples here studied by approximately 30% after each step. Cutaneous leishmaniasis (CL) is a vector-borne disease, which affects up to 1.5 million persons annually in tropical and subtropical regions worldwide (5). In several areas of endemicity like Peru, Leishmania species identification is important because different species with various clinical phenotypes coexist (11). There, higher incidence of mucocutaneous leishmaniasis (MCL) is attributed to Leishmania (Viannia) braziliensis (4), and low-pentavalent-antimonial treatment failures are associated with Leishmania (Viannia) guyanensis infections (1). Recognized associations between species and disease outcomes (1, 10, 12) have led the WHO expert committee on the control of leishmaniasis to recommend species identification for optimal treatment and control (16).PCR identification of Leishmania (Viannia) from clinical specimens (8, 14, 15) has used different targets: mannose phosphate isomerase (mpi) (17) and cysteine proteinase B (cpb) and the 70-kDa heat shock protein (hsp70) (6, 7, 9). These targets have been used successfully on CL specimens collected by filter paper lesion impressions (FPLIs) (3).Here, we report the development of a three-step strategy of PCR-based assays for Leishmania (Viannia) species identification and assess its performance on CL FPLIs. Reductions of time and cost to obtain results in areas where leishmaniasis is endemic were the criteria on which the algorithm was evaluated.DNA isolation from cultured parasites and filter paper lesion impressions. International reference strains and cultured parasites obtained from 80 patients attending our reference center were harvested, and their DNA was extracted as previously described (13). DNAs from anonymized, FPLI specimens from 70 CL lesions collected from 57 Peruvian patients were obtained as reported previously (2, 3). Codes and geographical origins are presented in Table S1 in the supplemental material.PCR assays for species identification. Leishmania (Viannia) species were identified using three PCR targets. The mpi gene was amplified in two separate reactions (mpi Lp and mpi Lb for Leishmania [Viannia] peruviana and L. [V.] braziliensis, respectively) according to the work of Zhang et al. (17). cpb and hsp70 genes were amplified as previously reported (7, 9). Primer sequences and reaction conditions for each target are described in Table S2 in the supplemental material. Analysis of cultured isolates and algorithm development.Eighty cultured isolates were analyzed using the three targets, of which 41 were identified as L. Table S1 in the supplemental material). Only 1 out of 80 remained unidentifiable after the three targets were assayed. Each one of the targets ident...

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“…26 They offer some advantages over biopsy techniques, including easy transportation from hospitals to diagnostic laboratories and the possibility of storing them for years when frozen and months when kept at room temperature. 11 In addition, filter papers are more likely to be favored by patients and clinics because they represent a non-invasive technique for collecting samples.…”

Section: Discussionmentioning

confidence: 99%

The Use of Fluorescent Fragment Length Analysis (PCR-FFL) in the Direct Diagnosis and Identification of Cutaneous Leishmania Species

Tomás-Pérez

Fisa

Riera

2013

The American Society of Tropical Medicine and Hygiene

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Abstract. Leishmaniasis is a disease caused by different species belonging to the genus Leishmania. It presents different epidemiological and clinical features and requires the development of rapid, sensitive techniques to improve specific diagnosis. In this study, we compared the traditional technique of polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) with PCR-fluorescent fragment length analysis (PCR-FFL). Fluorescently tagged primers, designed in the rRNA fragment ITS-1 and 7SL region, were used to amplify fragments, which were later digested and whose sizes were accurately determined using an automated DNA sequencer. We validated the technique using 19 Leishmania strains from five cutaneous Leishmania species before testing 36 clinical samples: 23 skin biopsies and 13 skin scrapings/lesion exudates on filter paper. In real diagnostic, PCR-FFL has proved to be quick, accurate, and more sensitive (83.3% testing the ITS-1 fragment and 94.4% testing the 7SL) than PCR-RFLP analysis (75% and 80.6%). Filter papers improved the specific diagnosis in both techniques using non-invasive samples.

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Diagnostic Performance of Filter Paper Lesion Impression PCR for Secondarily Infected Ulcers and Nonulcerative Lesions Caused by Cutaneous Leishmaniasis

Cited by 23 publications

References 21 publications

Simultaneous Infection with Leishmania (Viannia) braziliensis and L. (V.) lainsoni in a Peruvian Patient with Cutaneous Leishmaniasis

Simultaneous Infection with Leishmania (Viannia) braziliensis and L. (V.) lainsoni in a Peruvian Patient with Cutaneous Leishmaniasis

Leishmania (Viannia) Species Identification on Clinical Samples from Cutaneous Leishmaniasis Patients in Peru: Assessment of a Molecular Stepwise Approach

The Use of Fluorescent Fragment Length Analysis (PCR-FFL) in the Direct Diagnosis and Identification of Cutaneous Leishmania Species

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