Validation of an improved reference freeze-dried direct agglutination test for detecting leishmaniasis in the canine reservoir.

Access microbiology Pub Date : 2025-01-06 eCollection Date: 2025-01-01 DOI:10.1099/acmi.0.000890.v4
Abdallah El Harith, Elfadil Abass, Franjo Martinkovic, Durria Mansour, Hussam Ali Osman
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Abstract

Introduction. Proper identification and management of post-kala-azar dermal leishmaniasis (PKDL) and canine leishmaniasis (CanL) cases are among the prerequisites to the effective control of visceral leishmaniasis worldwide. Unlike PKDL, CanL still awaits effective improvement because of its cryptic nature, absence of Leishmania parasites in lesions or lymph nodes and not complete sensitivity of some diagnostic tools in use. Because of the need for certain skills and equipment, both the liquid direct agglutination test and freeze-dried direct agglutination test (FD-DAT) versions are, in comparison with the indirect immunofluorescence antibody test (IFAT) or enzyme-linked immunosorbent assay (ELISA), practical and feasible diagnostic alternatives. Aim. Validate the performance of an improved FD-DAT to suit routine and large-scale applications in CanL endemic areas. Methodology. Introducing citrate-saline formaldehyde (CSF) as an anti-clumping agent to replace normal saline for antigen reconstitution and drastically, however, eligibly lower the concentration of promastigotes (1.4×107) in comparison with the original FD-DAT reference (>5×107 ml-1). To ensure optimal safety, β-mercaptoethanol was replaced by urea or SDS as a serum-reducing agent. Results. By improving the procedure for reconstitution of FD-DAT antigen with CSF, a 150% reduction in the test application cost was achieved. Expired test batches (±4 years earlier) were successfully revitalized to full validity. As compared to the 48 h shelf-life time for the original, an FD-DAT batch reconstituted here with CSF maintained stability for ±12 months. Conclusions. The highly concordant results with IFAT and ELISA (one-way ANOVA test, P=0.142, homogeneity of variances P=0.009) as routine CanL diagnostics further motivate the application of the improved FD-DAT for the detection of the disease in endemic areas.

改进冻干直接凝集法检测犬库利什曼病的验证。
介绍。正确识别和管理黑热病后皮肤利什曼病(PKDL)和犬利什曼病(CanL)病例是世界范围内有效控制内脏利什曼病的先决条件之一。与PKDL不同,由于CanL的隐蔽性,病变或淋巴结中没有利什曼原虫,以及目前使用的一些诊断工具不完全敏感,CanL仍有待有效改进。由于需要一定的技能和设备,与间接免疫荧光抗体试验(IFAT)或酶联免疫吸附试验(ELISA)相比,液体直接凝集试验和冻干直接凝集试验(FD-DAT)版本都是实用可行的诊断替代方法。的目标。验证改进FD-DAT的性能,以适应CanL流行地区的常规和大规模应用。方法。引入柠檬酸盐-生理盐水甲醛(CSF)作为抗凝剂来代替生理盐水进行抗原重构,然而,与原始FD-DAT参考文献(>5×107 ml-1)相比,可以显著降低promastigotes的浓度(1.4×107)。为了确保最佳的安全性,用尿素或SDS代替β-巯基乙醇作为血清还原剂。结果。通过改进用CSF重组FD-DAT抗原的程序,测试应用成本降低了150%。过期的测试批次(±4年前)成功恢复到完全有效。与原始的48小时保质期相比,在CSF中重建的FD-DAT批次保持了±12个月的稳定性。结论。与IFAT和ELISA作为常规诊断方法的结果高度一致(单因素方差分析,P=0.142,方差齐性P=0.009),进一步促进了改良FD-DAT在流行地区的应用。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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