A PCR and RFLP-based molecular diagnostic algorithm for visceral leishmaniasis
作者机构：Laboratorio de Parasitologia MédicaDepartamento de Moléstias Infecciosas e Parasitárias do Hospital das Clinicas HCFMUSPFaculdade de MedicinaUniversidade de Sao PauloSao PauloBrazil Laboratorio de Imunopatologia e Biologia Molecular-Instituto Aggeu Magalhaes/IAM-FIOCRUZRecifeBrazil Laboratorio de Soroepidemiologia e Imunobiologia-FMUSPSao PauloBrazil Laboratorio de Biologia Molecular de Parasitas e Fungos-Instituto Adolfo LutzSao PauloBrazil Laboratorio de Parasitologia-Instituto de Medicina Tropical de Sao Paulo-USPSao PauloBrazil
出 版 物：《Asian Pacific Journal of Tropical Medicine》 (亚太热带医药杂志（英文版）)
年 卷 期：2020年第13卷第2期
页 面：62-70页
核心收录：
学科分类：100208[医学-临床检验诊断学] 1002[医学-临床医学] 100201[医学-内科学(含：心血管病、血液病、呼吸系病、消化系病、内分泌与代谢病、肾病、风湿病、传染病)] 10[医学]
基 金：supported by a grant from Fundacao de Amparo a Pesquisa no Estado de Sao Paulo(FAPESP)with grant number 2010-50304-8 under the supervision of Dr.Lucia Maria Almei Braz
主 题：Leishmania infantum Molecular diagnosis Visceral leishmaniasis PCR RFLP
摘 要：Objective:To determine an algorithm for molecular diagnosis of visceral leishmaniasis(VL)by kinetoplast DNA(kDNA)(RV1/RV2)and internal transcriber spacer(ITS1)(LITSR/L5.8 S)polymerase chain reaction(PCR),complemented by ITS 1 PCR restriction fragment length polymorphism(RFLP),using peripheral blood or bone marrow aspirate from patients with suspected ***:Biological samples were submitted to the gold standard for the diagnosis of VL and molecular diagnosis represented by ITS 1 PCR,kDNA PCR,and ITS 1 PCR *** samples were obtained from seven groups:groupⅠ,82 samples from patients with confirmed VL;groupⅡ,16 samples from patients under treatment for VL;groupⅢ,14 samples from dogs with canine visceral leishmaniasis(CVL);groupⅣ,a pool of six experimentally infected sandflies(Lutzomya longipalpis);group V,18 samples from patients with confirmed tegumentary leishmaniasis(TL)and groupsⅥandⅦwere from control groups without ***:The following gold standard and molecular examination results were obtained for each of the seven groups:groupⅠ:parasitologic and immunochromatographic tests showed a sensitivity of 76.3%(61 of 80)and 68.8%(55 of 80),respectively,and a sensitivity of 97.6%(80 of 82)and 92.7%(76 of 82)by ITS1 and kDNA PCR,*** ITS1 PCR RFLP(HaeⅢ)analysis of the 80 positive samples,52.5%(42 of 80)generated three fragments of 180,70,and 50 bp,corresponding to the pattern of Leishmania infantum infantum;groupⅡ:negative for the parasitologic methods and positive for IrK39(100%,16 of 16),presented 12.5%(2 of 16)of positivity by ITS 1 PCR and 25.0%(4 of 16)by kDNA PCR;groupⅢ:positive in the parasitologic and serologic tests(100%,14 of 14),presented 85.7%(12 of 14)of positivity by ITS1 PCR and kDNA ***1 PCR RFLP showed that 83.3%(10 of 12)of the canine samples contained parasites with profiles similar to ***;groupⅣpresented amplifications by ITS1 PCR and kDNA ***1 PCR products were analyzed by RFLP,generating a profile similar to
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