Polymerase chain reaction with lesion scrapping for the diagnosis of human American tegumentary leishmaniasis

Venazzi, Eneide Aparecida Sabaini; Roberto, Andréa Claudia Bekner Silva; Barbosa-Tessmann, Ione Parra; Zanzarini, Paulo Donizeti; Lonardoni, Maria Valdrinez Campana; Silveira, Thaís Gomes Verzignassi

doi:10.1590/s0074-02762006000400014

Cited by 19 publications

(23 citation statements)

References 18 publications

(30 reference statements)

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“…Some studies present molecular biology as a diagnostic tool for ACL with high sensitivity, generally giving positive findings in 100% of the patients analyzed as well as permitting the specific diagnosis of Leishmania 19 , 24 , 25 , 26 , 27 . However, it is, though, expensive and unavailable under the public health network of the State.…”

Section: Discussionmentioning

confidence: 99%

Epidemiological aspects of American cutaneous leishmaniasis (ACL) in an endemic area of forest extractivist culture in western Brazilian Amazonia

Brilhante

Melchior

Nunes

et al. 2017

Rev. Inst. Med. trop. S. Paulo

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The study sought to analyze clinical and epidemiological aspects of cutaneous leishmaniasis (CL) in Xapuri, Acre, Brazil. Data from 2008 to 2014 registered in the notification records of the disease of the Information System of the Complaints of Notification (SINAN), and the data of the Information Department of the Unified Health System (DATASUS) available from 2007 to 2013 were used and analyzed in the light of the statistics of the temporal series by the Prais-Winsten method and chi-squared test. A total of 906 cases were registered with 60.2% occurring in men and 39.7% in women. The groups from 0 to 4 years of age (48.0%) and from 5 to 19 years of age (23.3%) were the most affected. Regarding the clinical forms, 77.7% presented CL and 22.3% mucocutaneous leishmaniasis (MCL). Among the 896 cases with information on the diagnostic methods used, Montenegro’s skin test predominated (66.4%), with a positive result of 95.8% for CL and 99.3% for MCL. Treatment with N-methylglucamine antimony was performed in 99.4% of the cases, but discontinuously used in the majority of patients. This study presents information which may be used as a tool for the epidemiological surveillance and control of the disease in Xapuri, a region which depends essentially on forest resources and ecological tourism.

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Section: Discussionmentioning

confidence: 99%

Epidemiological aspects of American cutaneous leishmaniasis (ACL) in an endemic area of forest extractivist culture in western Brazilian Amazonia

Brilhante

Melchior

Nunes

et al. 2017

Rev. Inst. Med. trop. S. Paulo

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“…13,26 However, there is great variation in the sensitivity of PCR, especially in relation to the method of DNA extraction, the choice of the oligonucleotide primers, 25,27,28 the clinical samples used, and the length of infection. 16 In the present study, 26 samples from dogs (lesion scrapings, biopsy, and blood) and 47 samples from humans (lesion scrapings), from an endemic area of ACL caused by L. (V.) braziliensis, were tested by PCR using these five pairs of primers. PCR using the MP3H-MP1L primers, which amplify a fragment of the kDNA minicircle region of the subgenus Leishmania (Viannia), showed high positivity in human and canine lesions, detecting all samples that had positive parasite search.…”

Section: Discussionmentioning

confidence: 99%

“…The DNA was extracted from the blood by either the chloroform-phenol method 15 or the guanidine-phenol method. 16 The DNA sediment was then resuspended in 50 μL of TE buffer (10 mM TRIS; 1 mM EDTA; pH 8.0).…”

Section: Biological Samplesmentioning

confidence: 99%

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Comparison of different primes for PCR-based diagnosis of cutaneous leishmaniasis

Oliveira¹,

Lonardoni²,

Teodoro³

et al. 2011

Braz J Infect Dis

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Objective: The objective of this study was to analyze different primers that are commonly used in epidemiological studies for the detection of Leishmania DNA by PCR, and to compare them to the conventional direct parasite search for American cutaneous leishmaniasis (ACL) diagnosis. Material and methods: Five pairs of primers, four of them derived from Leishmania kDNA sequences (MP3H-MP1L; B1-B2; LBF1-LBR1; 13A-13B), and one derived from the SL RNA (mini-exon) gene repeat (LU5A-LB3C), reported previously, were used. Results: The MP3H-MP1L primers were the best at amplifying the DNA, detecting 2 fg of Leishmania spp. DNA. The 13A-13B primers presented the worst performance, detecting 512 x 10 3 fg of DNA. Conclusion: The wide variation in the analytical sensitivity of the primers used in the PCR, and the significant differences from the conventional method of ACL diagnosis found in this study, emphasize the importance of standardizing the PCR technique, analyzing sensitivity, and selecting suitable oligonucleotide primers.

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“…More recent methods for detection of the parasite such as immunohistochemistry (IHC) [7][8][9]13,[15][16][17] and polymerase chain reaction (PCR) 6,11,12,18 have been shown to be more sensitive and are beginning to be established in the laboratory routine as auxiliary or alternative methods. Considering this detection and regarding diagnostic performance parameters (sensitivity, e.g.…”

Section: Introductionmentioning

confidence: 99%

Immunoperoxidase technique using an anti-Leishmania (L.) chagasi hyperimmune serum in the diagnosis of culture-confirmed American tegumentary leishmaniasis

Quintella

Cuzzi

Madeira

et al. 2009

Rev. Inst. Med. trop. S. Paulo

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The present study reports the production of the rabbit anti-Leishmania (L.) chagasi hyperimmune serum, the standardization of the immunohistochemistry (IHC) technique and the evaluation of its employment in cutaneous leishmaniasis (CL) lesions diagnosed by Leishmania sp. culture isolation. Thirty fragments of active CL lesions were examined as well as 10 fragments of cutaneous mycosis lesions as control group. IHC proved more sensitive in detecting amastigotes than conventional hematoxylin-eosin (HE) stained slides: the former was positive in 24 (80%) biopsies whereas the latter, in 16 (53%) (p = 0.028). The reaction stained different fungus species causing cutaneous mycosis. Besides, positive reaction was noticed in mononuclear and endothelial cells. Nevertheless, this finding was present in the control group biopsies. It is concluded that IHC showed good sensitivity in detecting amastigotes.

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Polymerase chain reaction with lesion scrapping for the diagnosis of human American tegumentary leishmaniasis

Abstract: The objective of this work was to compare the polymerase chain reaction (PCR)

Cited by 19 publications

References 18 publications

Epidemiological aspects of American cutaneous leishmaniasis (ACL) in an endemic area of forest extractivist culture in western Brazilian Amazonia

Epidemiological aspects of American cutaneous leishmaniasis (ACL) in an endemic area of forest extractivist culture in western Brazilian Amazonia

Comparison of different primes for PCR-based diagnosis of cutaneous leishmaniasis

Immunoperoxidase technique using an anti-Leishmania (L.) chagasi hyperimmune serum in the diagnosis of culture-confirmed American tegumentary leishmaniasis

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