聚合酶链反应（PCR）和寡核苷酸连接分析（NASBA）检测后应用寡色氨酸层析法进行布鲁氏菌和利什曼原虫的分子诊断

Accordance and concordance of PCR and NASBA followed by oligochromatography for the molecular diagnosis of Trypanosoma brucei and Leishmania.

机构信息

Department of Veterinary Parasitology and Microbiology, Makerere University, Kampala, Uganda.

出版信息

Trop Med Int Health. 2010 Jul;15(7):800-5. doi: 10.1111/j.1365-3156.2010.02547.x. Epub 2010 May 14.

DOI:10.1111/j.1365-3156.2010.02547.x

PMID:20487429

Abstract

OBJECTIVE

To evaluate the repeatability and reproducibility of four simplified molecular assays for the diagnosis of Trypanosoma brucei spp. or Leishmania ssp. in a multicentre ring trial with seven participating laboratories.

METHODS

The tests are based on PCR or NASBA amplification of the parasites nucleic acids followed by rapid read-out by oligochromatographic dipstick (PCR-OC and NASBA-OC).

RESULTS

On purified nucleic acid specimens, the repeatability and reproducibility of the tests were Tryp-PRC-OC, 91.7% and 95.5%; Tryp-NASBA-OC, 95.8% and 100%; Leish-PCR-OC, 95.9% and 98.1%; Leish-NASBA-OC, 92.3% and 98.2%. On blood specimens spiked with parasites, the repeatability and reproducibility of the tests were Tryp-PRC-OC, 78.4% and 86.6%; Tryp-NASBA-OC, 81.5% and 89.0%; Leish-PCR-OC, 87.1% and 91.7%; Leish-NASBA-OC, 74.8% and 86.2%.

CONCLUSION

As repeatability and reproducibility of the tests were satisfactory, further phase II and III evaluations in clinical and population specimens from disease endemic countries are justified.

摘要

目的

在一项有 7 个参与实验室的多中心环试验中，评估 4 种简化分子检测方法对诊断布氏锥虫 spp.或利什曼原虫 spp.的重复性和重现性。

方法

这些检测方法基于寄生虫核酸的 PCR 或 NASBA 扩增，随后通过寡核苷酸条带（PCR-OC 和 NASBA-OC）快速读取。

结果

在纯化的核酸标本上，检测方法的重复性和重现性分别为 Tryp-PRC-OC，91.7%和 95.5%；Tryp-NASBA-OC，95.8%和 100%；Leish-PCR-OC，95.9%和 98.1%；Leish-NASBA-OC，92.3%和 98.2%。在含有寄生虫的血液标本上，检测方法的重复性和重现性分别为 Tryp-PRC-OC，78.4%和 86.6%；Tryp-NASBA-OC，81.5%和 89.0%；Leish-PCR-OC，87.1%和 91.7%；Leish-NASBA-OC，74.8%和 86.2%。