Competitive Polymerase Chain Reaction Used to Diagnose Cutaneous Leishmaniasis in German Soldiers Infected During Military Exercises in French Guiana

Bezold, Guntram; Lange, Monika; Gethöffer, Kerstin; Pillekamp, H.; Reindl, Herrmann; Richter, C.; Schönian, Gabriele; Weber, Lothar W.; Ru, Peter

doi:10.1007/s100960100519

Cited by 6 publications

(6 citation statements)

References 8 publications

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“…In nine cases Leishmania was not identified microscopically; histopathological diagnoses were: granulomatous dermatitis (6), lupoid rosacea (1), foreign body granuloma (1) and granuloma annulare (1). Unaltered epidermis (9), nodular infiltrates (5), numerous multinucleated histiocytes (3), palisaded granulomas with fibrinoid centres (2), sarcoidal granulomas (4) and elastophagocytosis (1) misled the histopathologists in these cases. Conclusions CL seems often to be misdiagnosed clinically in countries where it is not endemic.…”

Section: Discussionmentioning

confidence: 99%

Unusual histopathological features of cutaneous leishmaniasis identified by polymerase chain reaction specific for Leishmania on paraffin-embedded skin biopsies

Böer

Blödorn‐Schlicht

Wiebels

et al. 2006

British Journal of Dermatology

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CL seems often to be misdiagnosed clinically in countries where it is not endemic. Histopathologically, CL may be misinterpreted as sarcoidosis, foreign body granuloma, lupoid rosacea and granuloma annulare, especially when Leishmania is not seen microscopically. We suggest that in Northern Europe, PCR for Leishmania-specific DNA should be performed routinely in any granulomatous dermatitis presenting as a single or few nodules on the extremities or face, even when a diagnosis of CL was not considered by the referring clinician.

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Section: Discussionmentioning

confidence: 99%

Unusual histopathological features of cutaneous leishmaniasis identified by polymerase chain reaction specific for Leishmania on paraffin-embedded skin biopsies

Böer

Blödorn‐Schlicht

Wiebels

et al. 2006

British Journal of Dermatology

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“…Also, logistical problems would be associated with its application in epidemiological dog surveys (e.g., dogs would have to be kenneled or monitored for 48 h prior reading the MST induration size). PCR has been shown to be particularly useful for the diagnosis of Leishmania (Viannia) infection, as parasite numbers in clinical samples are typically sparse (4,6,9,17,21,27,29,33,34,40,41).…”

Section: Discussionmentioning

confidence: 99%

“…As is the case for zoonotic visceral leishmaniasis (3,5,22,32,36), PCR on blood alone does not appear to be the elusive gold standard for diagnosing ACL infections in dogs (or humans). Unless a more sensitive PCR protocol (e.g., PCR-ELISA or different PCR primers) is developed to detect asymptomatic ACL infections, the use of PCR should be restricted to the diagnosis of active cases (e.g., hospitals in Europe, the United States, and elsewhere increasingly rely on PCR for Leishmania diagnosis in human patients) (6,41) and, in particular, MCL cases where common diagnostic tests (e.g., parasite culture, biopsy smears, and histopathology) are less sensitive. Nonetheless, the use of PCR on blood will have an important epidemiological application in studies monitoring the clinical and chemotherapeutic follow-up of ACL patients (10,15,34).…”

Section: Discussionmentioning

confidence: 99%

“…Various PCR protocols have been developed for the detection of ACL-causing Leishmania in humans by using either purified DNA (from cultured parasites) (9) or clinical specimens (including lesion and scar biopsy specimens [4,6,9,17,21,27,29,33,34,40] or blood [10,12,14,15,26]) and have consistently been proven to be more sensitive than other diagnostic methods, including in vitro culture of biopsy specimens, biopsy smears, and hamster inoculation. However, at present only one large (Ͼ50 samples tested) study has been reported where PCR was used as a diagnostic massscreening tool to detect ACL in putative animal reservoirs (20).…”

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confidence: 99%

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Evaluation of PCR as a Diagnostic Mass-Screening Tool To Detect Leishmania ( Viannia ) spp. in Domestic Dogs ( Canis familiaris )

et al. 2003

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Several studies have suggested that the PCR could be used in epidemiological mass-screening surveys to detect Leishmania (Viannia) spp. infection in human and animal hosts. Dogs from an area of Leishmania braziliensis and Leishmania peruviana endemicity were screened for American cutaneous leishmaniasis (ACL) infection by established PCR-based and enzyme-linked immunosorbent antibody test (ELISA) protocols. PCR detected Leishmania (Viannia) infection in a total of 90 of 1,066 (8.4%) dogs: 32 of 368 (8.7%), 65 of 769 (8.5%), and 7 of 42 (16.7%) dogs were PCR positive by testing of whole blood, buffy coat, and bone marrow aspirates, respectively. ELISA detected infection in 221 of 1,059 (20.9%) tested dogs. The high prevalence of Leishmania (Viannia) detected by PCR and ELISA in both asymptomatic (7.5 and 19.2%, respectively) and symptomatic (32 and 62.5%, respectively) dogs is further circumstantial evidence for their suspected role as reservoir hosts of ACL. However, the low sensitivity of PCR (31%) compared to ELISA (81%) indicates that PCR cannot be used for mass screening of samples in ACL epidemiological studies. Unless more-sensitive PCR protocols were to be developed, its use should be restricted to the diagnosis of active (canine and human) cases and to the parasitological monitoring of patients after chemotherapy.

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“…Numerous DNA-based assays, e.g. PCR plus sequencing, and restriction fragment length polymorphism (RFLP) analysis, have been developed for this purpose (Bezold et al, 2001;Mauricio et al, 2004;Chargui et al, 2005;Oliveira et al, 2005;Schönian et al, 2008). However, conventional PCR assays provide results in one or two working days (Bretagne, 2003;Foulet et al, 2007).…”

Section: Introductionmentioning

confidence: 99%

Diagnosis and identification ofLeishmaniaspp. from Giemsa-stained slides, by real-time PCR and melting curve analysis in south-west of Iran

Khademvatan

Neisi

Maraghi

et al. 2011

Annals of Tropical Medicine & Parasitology

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Background:The aim of present study was describing a real-time PCR assay for the diagnosis and direct identification of Leishmania species on Giemsa-stained slides in south-west of Iran.Materials and methods: Altogether, 102 Giemsa-stained slides were collected from different part of south-west of Iran between 2008 and 2011. All the Giemsa-stained slides were examined under light microscope. After DNA extraction, real-time PCR amplification and detection were conducted with fluorescent SYBR Green I. For identification, PCR products were analysed with melting curve analysis.Results: One hundred and two archived slides from suspected lesion examined by microscopy and real-time PCR. The sensitivity of the real-time PCR on Giemsa-stained slid was 98% (96/102). The melting curve analysis (T m ) were 88.3¡0.2uC for L. tropica (MHOM/IR/02/Mash10), 86.5¡0.2uC for L. major (MHOM/IR/75/ER) and 89.4¡0.3uC for L. infantum (MCAN/IR/97/LON 49), respectively.Conclusion: This study is first report in use of real-time PCR for diagnosis and identification of Leishmania spp. in Iran. Up to now, in Iran, the majority of identification of Leishmania species is restriction fragment length polymorphism (PCR-RFLP) of ITS1 and kinetoplast DNA. Our data showed that Giemsa-stained slides that were stored more than 3 years, can be use for Leishmania DNA extraction and amplification by real-time PCR. Compared to conventional PCR-based methods, the real-time PCR is extremely rapid with results and more samples can be processed at one time.

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Competitive Polymerase Chain Reaction Used to Diagnose Cutaneous Leishmaniasis in German Soldiers Infected During Military Exercises in French Guiana

Cited by 6 publications

References 8 publications

Unusual histopathological features of cutaneous leishmaniasis identified by polymerase chain reaction specific for Leishmania on paraffin-embedded skin biopsies

Unusual histopathological features of cutaneous leishmaniasis identified by polymerase chain reaction specific for Leishmania on paraffin-embedded skin biopsies

Evaluation of PCR as a Diagnostic Mass-Screening Tool To Detect Leishmania ( Viannia ) spp. in Domestic Dogs ( Canis familiaris )

Diagnosis and identification ofLeishmaniaspp. from Giemsa-stained slides, by real-time PCR and melting curve analysis in south-west of Iran

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