抗Sj29单克隆抗体的研制及其在血吸虫病诊断中的可能应用。

Development of monoclonal antibodies against Sj29 and its possible application for schistosomiasis diagnosis.

作者信息

Ren Cui-Ping, Liu Quan, Liu Feng-Chun, Zhu Feng-Yu, Cui Shi-Xiang, Liu Zhen, Gao Wen-Da, Liu Miao, Ji Yong-Sheng, Shen Ji-Jia

机构信息

Department of Microbiology and Parasitology, Anhui Provincial Laboratory of Pathogen Biology, Anhui Provincial Laboratory of Zoonoses, Laboratory of Tropical and Parasitic Diseases Control, School of Basic Medical Sciences, Anhui Medical University, Hefei, China; Department of Immunology, Anhui Medical University, Hefei, China.

出版信息

Int J Infect Dis. 2017 Aug;61:74-78. doi: 10.1016/j.ijid.2017.04.009. Epub 2017 Apr 12.

DOI:10.1016/j.ijid.2017.04.009

PMID:28412409

Abstract

OBJECTIVE

Timely Schistosoma japonicum detection improves outcomes in schistosomiasis. Here, we established a double antibody sandwich ELISA to detect Schistosoma japonicum.

METHODS

Sj29 polyclonal and monoclonal antibodies were developed and identified. A Sj29 double antibody sandwich ELISA was evaluated.

RESULTS

Assay sensitivity for detecting Schistosoma japonicum circulating antigen Sj29 was 76.7% (23/30), 54.5% (18/33) and 50.0% (18/36) in patients with acute, chronic and advanced schistosomiasis. No false positives or cross-reactivity was observed in healthy controls or patients with clonorchiasis, paragonimiasis, or ancylostomiasis, respectively. By contrast, false positives (5.7%) and cross-reactivity (6.5%-10%) were detected using an AWA-ELISA. The circulating antigen positive rates decreased significantly faster than that of the antibody detection after 6 months treatment (22.2%, 4/18 and 88.9%, 16/18). Chi-Square Tests revealed that Sj29 sandwich ELISA had lower sensitivity than AWA indirect ELISA in the detection of S. japonicum infected patients (p<0.05). Although our assay detection specificity in patients infected with other parasites or healthy controls appeared higher, the difference between the assays was insignificant. However, our assay showed significantly better results in monitoring praziquantel therapeutic effects (p=0.001), with antigen-positive rates decreasing significantly faster than antibody detection rates after 6 months of treatment (22.2%, 4/18 versus 88.9%, 16/18).

CONCLUSIONS

Sj29 double antibody sandwich ELISA was established. The specificity of this method for detecting healthy sera was 100%. Meanwhile, Sj29 sandwich ELISA may have a potential diagnostic capability to distinguish current from past infections and assess drug treatment responses.

摘要

目的

及时检测日本血吸虫可改善血吸虫病的治疗效果。在此，我们建立了一种双抗体夹心ELISA法来检测日本血吸虫。

方法

制备并鉴定了Sj29多克隆抗体和单克隆抗体。对Sj29双抗体夹心ELISA法进行了评估。

结果

检测日本血吸虫循环抗原Sj29的检测灵敏度在急性、慢性和晚期血吸虫病患者中分别为76.7%（23/30）、54.5%（18/33）和50.0%（18/36）。在健康对照者或华支睾吸虫病、肺吸虫病或钩虫病患者中未观察到假阳性或交叉反应。相比之下，使用AWA-ELISA检测到假阳性（5.7%）和交叉反应（6.5%-10%）。治疗6个月后，循环抗原阳性率下降明显快于抗体检测阳性率（22.2%，4/18和88.9%，16/18）。卡方检验显示，Sj29夹心ELISA法在检测日本血吸虫感染患者时的灵敏度低于AWA间接ELISA法（p<0.05）。尽管我们的检测方法在感染其他寄生虫的患者或健康对照者中的检测特异性似乎更高，但两种检测方法之间的差异不显著。然而，我们的检测方法在监测吡喹酮治疗效果方面显示出明显更好的结果（p=0.001），治疗6个月后抗原阳性率下降明显快于抗体检测率（22.2%，4/18对88.9%，16/18）。