Performance of parasitological and molecular techniques for the diagnosis and surveillance of gambiense sleeping sickness.
Performance of parasitological and molecular techniques for the diagnosis and surveillance of gambiense sleeping sickness.
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DOI：
10.1371/journal.pntd.0002954
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发表时间：
2014-06
影响因子：
3.8
通讯作者：
Lejon V
中科院分区：
文献类型：
--
作者：
Mumba Ngoyi D;Ali Ekangu R;Mumvemba Kodi MF;Pyana PP;Balharbi F;Decq M;Kande Betu V;Van der Veken W;Sese C;Menten J;Büscher P;Lejon V
最近，在控制冈比亚昏睡病的诊断方面取得了改进，但其表现仍然缺乏记录，可能取决于检查前的标本处理。在刚果民主共和国的一项前瞻性研究中，我们比较了几种寄生虫检测技术、免疫锥虫酶解和m18S PCR对储存在稳定缓冲液中或用滤纸干燥的全血的诊断性能。对CATT全血（WB）滴度≥1∶4或有昏睡病临床体征的患者，检查淋巴结（LNA）和（或）血液中是否存在锥虫。采用毛细管离心法（CTC）、微阴离子交换离心法（mAECT）和白毛离心法（BC）检测血液。全血(i)保存在盐酸胍EDTA （GE）稳定缓冲液中，（ii）在滤纸上干燥，进行PCR，并评估重复性和再现性。血浆进行免疫锥虫酶解（TL）。共包括237人。143例寄生虫学确诊病例中，85.3%的ct - wb滴度≥1/8，LNA阳性39.2%，CTC阳性47.5%，mAECT-WB阳性80.4%，mAECT-BC阳性90.9%，TL阳性95.1%，ge稳定血PCR阳性高达89.5%。ge稳定血PCR结果重复性最高（87.8%），实验室间重复性最高（86.9%）。94例未确诊病例中，TL或PCR阳性分别为39.4%和23.4%。潜类分析也提示PCR和TL的特异性不佳。LNA检查与mAECT-BC结合提供了极好的诊断敏感性。对于PCR，将血液储存在稳定缓冲液中优于滤纸。TL和PCR对远程诊断是有用的，但并不比mAECT-BC更敏感。对于TL和PCR，其特异性及其对未确诊疑似病例管理的有用性仍有待确定。非洲人类锥虫病或昏睡病仍在撒哈拉以南非洲造成相当大的痛苦。这种传染病的诊断方法不断改进，但在控制活动中实施之前，应评估其准确性和可重复性方面的表现。我们评估了几种显微镜、血清学和分子诊断测试对刚果民主共和国237名昏睡病疑似患者的诊断性能。由于分子诊断相当复杂，我们还评估了它们的可重复性和再现性。在缺乏金标准测试的情况下，潜在类分析显示，血清学和分子测试的次优特异性是一个问题。本研究表明，结合淋巴结抽吸检查和微型阴离子交换离心技术从血液中分离锥虫具有较高的诊断敏感性。
Recently, improvements have been made to diagnostics for gambiense sleeping sickness control but their performance remains poorly documented and may depend on specimen processing prior to examination. In a prospective study in the Democratic Republic of the Congo, we compared the diagnostic performance of several parasite detection techniques, immune trypanolysis and of m18S PCR on whole blood stored in a stabilisation buffer or dried on filter paper. Individuals with CATT whole blood (WB) titer ≥1∶4 or with clinical signs indicative for sleeping sickness were examined for presence of trypanosomes in lymph node aspirate (LNA) and/or in blood. Blood was examined with Capillary Centrifugation Technique (CTC), mini-Anion Exchange Centrifugation Technique (mAECT) and mAECT on buffy coat (BC). PCR was performed on whole blood (i) stored in guanidine hydrochloride EDTA (GE) stabilisation buffer and (ii) dried on filter paper, and repeatability and reproducibility were assessed. Immune trypanolysis (TL) was performed on plasma. A total of 237 persons were included. Among 143 parasitologically confirmed cases, 85.3% had a CATT-WB titre of ≥1/8, 39.2% were positive in LNA, 47.5% in CTC, 80.4% in mAECT-WB, 90.9% in mAECT-BC, 95.1% in TL and up to 89.5% in PCR on GE-stabilised blood. PCR on GE-stabilised blood showed highest repeatability (87.8%) and inter-laboratory reproducibility (86.9%). Of the 94 non-confirmed suspects, respectively 39.4% and 23.4% were TL or PCR positive. Suboptimal specificity of PCR and TL was also suggested by latent class analysis. The combination of LNA examination with mAECT-BC offered excellent diagnostic sensitivity. For PCR, storage of blood in stabilisation buffer is to be preferred over filter paper. TL as well as PCR are useful for remote diagnosis but are not more sensitive than mAECT-BC. For TL and PCR, the specificity, and thus usefulness for management of non-confirmed suspects remain to be determined. Human African trypanosomiasis or sleeping sickness still causes considerable suffering in sub-Sahara Africa. Diagnostics for this infectious disease constantly improve but their performance in terms of accuracy and reproducibility should be evaluated prior to implementation in control activities. We evaluated the diagnostic performance of several microscopic, serological and molecular diagnostic tests on a cohort of 237 sleeping sickness suspects in the Democratic Republic of the Congo. Since molecular diagnostics are rather sophisticated, we also assessed their repeatability and reproducibility. In the absence of a golden standard test, latent class analysis revealed that the suboptimal specificity of the serological and molecular tests is an issue. Our study shows the superior diagnostic sensitivity of the combination of lymph node aspirate examination and separation of trypanosomes from blood by mini Anion Exchange Centrifugation Techniques.
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影响因子：
3.8
作者：
Jamonneau V;Ilboudo H;Kaboré J;Kaba D;Koffi M;Solano P;Garcia A;Courtin D;Laveissière C;Lingue K;Büscher P;Bucheton B
通讯作者：
Bucheton B
影响因子：
2.1
作者：
LANHAM, SM;GODFREY, DG
通讯作者：
GODFREY, DG
影响因子：
2.7
作者：
Koffi, M.;Solano, P.;Jamonneau, V.
通讯作者：
Jamonneau, V.
影响因子：
3.8
作者：
Deborggraeve, Stijn;Lejon, Veerle;Buscher, Philippe
通讯作者：
Buscher, Philippe
影响因子：
3.8
作者：
Buescher, Philippe;Ngoyi, Dieudonne Mumba;Bieler, Sylvain
通讯作者：
Bieler, Sylvain