Detection of cutaneous Leishmania infection in paraffin-embedded skin biopsies using the polymerase chain reaction

Laskay, Tamás; Mikó, T; Negesse, Yohannes; Solbach, Werner; Röllinghoff, Martin; Frommel, D.

doi:10.1016/0035-9203(95)90537-5

Cited by 75 publications

(78 citation statements)

References 17 publications

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“…Of all molecular methods used for species determination, PCR-RFLP is proved to be the most sensitive and specific technique (de Oliveira et al, 2003;Culha et al, 2006). Skin biopsy and also materials obtained by skin scrape/exudate have been used for PCR in different studies (Laskay et al, 1995). The objective of this study was therefore to identify and molecular characterize the parasites responsible for CL in endemic area of Yazd, Iran, using PCR-RFLP.…”

Section: Introductionmentioning

confidence: 99%

The changing profile of cutaneous leishmaniasis agent in a central province of Iran

Eslami

Anvarii

Ebadi

et al. 2013

Tanzania J Hlth Res

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Cutaneous leishmaniasis in Iran is usually caused by Leishmania major or L. tropica. However, the direct examination or the cultures of biopsies for diagnosis are not very sensitive. The objective of this study was to identify the responsible species obtained from patients suspected of cutaneous leishmaniasis referred to the reference laboratory at Yazd in Iran during 2010-2011 using parasitological and molecular assays. After completing a clinical/epidemiologic data questionnaire for 145 patients with suspected skin lesions, scraping samples were collected. Each specimen was examined using both direct microscopy and molecular assay using polymerase chain reaction-restriction length polymorphism (PCR-RFLP). Location of the lesions included 47.7% on hands, 30.7% on face, 15.4% on feet, and the remainder on other regions. Out of 145 samples, Leishman body was observed in 52 by direct smear and 73 by PCR assay. Molecular assay indicated 36 cases as L. major, 36 cases as L. tropica and one case as unknown. In conclusion, molecular characterization showed changing profile of Leishmania species in the study area which may have implications on treatment and/or control strategies. ______________________________________________________________________________________

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Section: Introductionmentioning

confidence: 99%

The changing profile of cutaneous leishmaniasis agent in a central province of Iran

Eslami

Anvarii

Ebadi

et al. 2013

Tanzania J Hlth Res

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“…However, FFPE samples can be stored without cooling, which facilitates their transportation and storage in locations with poor infrastructure (Laskay et al, 1995). In this study, we evaluated the accuracy of quantitative polymerase chain reaction (qPCR) conducted with FT and FFPE samples of healthy skin, collected from the scapular region of dogs, for L. infantum DNA detection.…”

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confidence: 99%

Accuracy of quantitative polymerase chain reaction in samples of frozen and paraffin-embedded healthy skin for the diagnosis of canine visceral leishmaniasis

Campos

Madeira

Silva³

et al. 2017

Arq. Bras. Med. Vet. Zootec.

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“…No false positive results were obtained. Laskay et al (1995), using oligonucleotide primers 13A and 13B were able to detect L. aethiopica in paraffin-embedded tissue, in 7 out of the 22 cases where the conventional methods failed. They proposed the PCR as a valid diagnostic method for leishmaniasis, especially for chronic cases of the pathology.…”

Section: Discussionmentioning

confidence: 96%

Detection of Leishmania braziliensis in human paraffin-embedded tissues from Tucumán, Argentina by polymerase chain reaction

Córdoba-Lanús

Piñero

González

et al. 2005

Mem. Inst. Oswaldo Cruz

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American cutaneous leishmaniasis (ACL) is an endemic disease in Northern Key words: Leishmania (V.) braziliensis -diagnosis -polymerase chain reaction -ArgentinaAmerican tegumentary leishmaniasis (cutaneous and mucocutaneous) has been endemic in Argentina since 1916(Mazza 1926. In the Northern provinces of the country, outbreaks were identified in 1985/1987, 1997/1998. In Tucumán province, between 1991and 1996 leishmaniasis cases were reported (Marcolongo et al. 1993, Yadón 1997, Villalonga 1998.In the Americas the causal agents of this pathology are the mexicana complex species: and L. (L.) in the country (Sosa-Estani et al. 1998). At present, the characterization of parasites from human patients in Tucumán has not been possible. Studies from epidemic outbreaks in nearby provinces within Argentina, indicate that the parasites belong to the L. braziliensis complex (Campanini et al. 1993, Cuba et al. 1996, Sosa-Estani et al. 1998. The presence of L. (L.) amazonensis (L. mexicana complex) by isoenzymatic analysis has also been reported in the Chaco region, Argentina (Frank et al. 2000).The most common diagnostic method for cutaneous leishmaniasis used in Argentina is the smear -histopathology exam in combination with clinical and epidemiological data. Mucocutaneous leishmaniasis generally requires a complementary test because of the scarcity of parasites in the lesions.The polymerase chain reaction technique (PCR) has been successfully employed in the detection of specific sequences of pathogenic agents (Erlich 1991). At present this technique is an adequate method for the leishmaniasis diagnosis, showing a high sensitivity when compared to traditional tests (Rodgers et al. 1990, De Bruijn & Barker 1992, De Bruyin et al. 1993, Lopez et al. 1993a, b, Meredith et al. 1993, Piarroux et al. 1994. The sensitivity of the laboratory diagnosis increases significantly when PCR is combined with Southern blotting (Andresen et al. 1996).In the present study we used PCR and Southern blotting for the detection of Leishmania spp. and identification of the parasite from skin biopsies, histologically examined, of human patients from an endemic area of Tucumán province, Argentina. MATERIALS AND METHODSSamples -In this study we used 22 paraffin-embedded skin biopsies from endemic areas of leishmaniasis in the province of Tucumán, kindly donated by the Pathologic Anatomy Service of the Angel C Padilla Hospital (Tucumán, Argentina). Twenty one of the samples had a clinical diagnosis for leishmaniasis and the histopathology test was positive for 13 of them and negative for the rest (Table I). The only sample with a nonleishmanial clinical diagnosis and a negative histopathology result was that used as a negative control.DNA extraction -For each sample five to seven 5 µm sections of paraffin-embedded tissue were placed in a tube. The samples were washed with xilene, incubated for 30 min at 37ºC, centrifuged at 13,000 rpm for 5 min and the supernatant discarded. The washing procedure was repeated three times. Then two washes with eth...

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Detection of cutaneous Leishmania infection in paraffin-embedded skin biopsies using the polymerase chain reaction

Cited by 75 publications

References 17 publications

The changing profile of cutaneous leishmaniasis agent in a central province of Iran

The changing profile of cutaneous leishmaniasis agent in a central province of Iran

Accuracy of quantitative polymerase chain reaction in samples of frozen and paraffin-embedded healthy skin for the diagnosis of canine visceral leishmaniasis

Detection of Leishmania braziliensis in human paraffin-embedded tissues from Tucumán, Argentina by polymerase chain reaction

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