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酶联免疫电转移印迹法进一步评价重组 Tsol-p27 对莫桑比克猪囊尾蚴病血清学诊断的价值。

Further evaluation of recombinant Tsol-p27 by enzyme-linked immunoelectrotransfer blot for the serodiagnosis of cysticercosis in pigs from Mozambique.

机构信息

Department of Microbiology, Faculty of Medicine, Eduardo Mondlane University, Maputo, Mozambique.

Department of Cell and Molecular Biology, BMC, Uppsala University, Uppsala, Sweden.

出版信息

Parasit Vectors. 2019 Nov 27;12(1):564. doi: 10.1186/s13071-019-3816-x.

Abstract

BACKGROUND

Porcine cysticercosis has a negative impact on human health and the meat industry, as it makes infected meat unaproprieted for consuption and it is the main etiology of epileptic seizures in developing countries. There are multiple serological assays that use crude antigens with high sensitivity and specificity for the diagnosis of both porcine and human cysticercosis. Nonetheless, antigen preparation is time-consuming, needs a well-equipped laboratory and trained personnel and places those manipulating the meat at great risk for infection. New serodiagnostic approaches to the diagnosis of porcine and human cysticercosis have been directed towards the development of recombinant deoxyribonucleic acid technology for the generation of synthetic proteins that can serve as simplified, low-cost and harmless substitutes for native antigens. The aim of the present study was to further evaluate the recombinant Tsol-p27 protein as a target molecule in immunoassays for the serodiagnosis of porcine cysticercosis. From these data, we hoped to develop recommendations regarding its use in the serodiagnosis of porcine cysticercosis.

RESULTS

We studied a panel of 83 naturally infected pig sera from Angónia District, Mozambique, an endemic area for porcine and human cysticercosis. These sera were previously tested by antigen enzyme-linked immunosorbent assay (Ag-ELISA) to detect antigens of T. solium. The serum panel was processed by enzyme-linked immunoelectrotransfer blot (EITB) assay against the recombinant Tsol-p27 protein and the Ag-ELISA assay results were used to compare and evaluate the performance of Tsol-p27 for the diagnosis of cysticercosis. Out of 83 sera, 24 (29.0%) were positive for Tsol-p27 and 59 (71%) were negative in the same assay. From the 37 sera that tested positive to Ag-ELISA, 11 (13.3%) were positive to Tsol-p27, while from 46 sera that tested negative to Ag-ELISA, 33 (39.7%) also tested negative to Tsol-p27. The sensitivity and specificity of Tsol-p27 was 29.7% and 71.7%, respectively, while the positive predictive value and negative predictive value were 45.8% and 55.9%, respectively, as calculated using Medcalc® version 15.0 software (MedCalc Software, Ostend, Belgium).

CONCLUSION

While Tsol-p27 recombinant protein might be suitable for testing human sera, its performance in pigs is not acceptable, so other recombinant proteins should be evaluated alone or multiplexed.

摘要

背景

猪囊尾蚴病对人类健康和肉类行业有负面影响,因为它使受感染的肉不适合食用,而且是发展中国家癫痫发作的主要病因。有多种血清学检测方法使用粗抗原,具有高灵敏度和特异性,可用于诊断猪和人类囊尾蚴病。然而,抗原制备耗时,需要设备齐全的实验室和训练有素的人员,并且使处理肉的人员面临很大的感染风险。针对猪和人类囊尾蚴病的新血清学诊断方法已针对脱氧核糖核酸重组技术的发展,用于生成可作为天然抗原简化、低成本和无害替代品的合成蛋白。本研究的目的是进一步评估重组 Tsol-p27 蛋白作为免疫分析中用于诊断猪囊尾蚴病的靶分子。根据这些数据,我们希望就其在猪囊尾蚴病血清学诊断中的应用提出建议。

结果

我们研究了来自莫桑比克 Angónia 区的 83 份自然感染猪血清,该地区是猪和人类囊尾蚴病的流行区。这些血清先前已通过抗原酶联免疫吸附试验(Ag-ELISA)检测了猪带绦虫抗原。用酶联免疫电泳转移印迹(EITB)试验对重组 Tsol-p27 蛋白处理血清组,并将 Ag-ELISA 试验结果用于比较和评估 Tsol-p27 对囊尾蚴病的诊断性能。在 83 份血清中,24 份(29.0%)对 Tsol-p27 呈阳性,在同一检测中 59 份(71%)呈阴性。在对 Ag-ELISA 呈阳性的 37 份血清中,11 份(13.3%)对 Tsol-p27 呈阳性,而在对 Ag-ELISA 呈阴性的 46 份血清中,33 份(39.7%)也对 Tsol-p27 呈阴性。Tsol-p27 的灵敏度和特异性分别为 29.7%和 71.7%,阳性预测值和阴性预测值分别为 45.8%和 55.9%,使用 Medcalc®版本 15.0 软件(MedCalc Software,Ostend,比利时)计算得出。

结论

虽然 Tsol-p27 重组蛋白可能适合测试人类血清,但在猪中的性能不可接受,因此应单独或多重评估其他重组蛋白。

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