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[用于快速检测创伤弧菌的免疫层析试纸条检测法的研发]

[Development of an immunochromatographic strip test for rapid detection of Vibrio vulnificus].

作者信息

Yan Zhi-min, Zheng Jing, Chen Qing, Shen Hong

机构信息

Department of Pathology, Southern Medical University, Guangzhou, China.

出版信息

Nan Fang Yi Ke Da Xue Xue Bao. 2011 May;31(5):894-6.

Abstract

OBJECTIVE

To establish a rapid strip test for detection of Vibrio vulnificus.

METHOD

Anti-Vibrio vulnificus polyclonal antibodies were obtained from the rabbits immunized with Vibrio vulnificus (ATCC27562). Colloidal gold was prepared through reducing HAuCl(4)·4H(2)O by sodium citrate and conjugated with the polyclonal antibodies as the detecting reagent. The polyclonal antibodies and sheep anti-rabbit immunoglobulins were separately coated onto the same nitrocellulose membrane for sample detection and quality-control, respectively. The nitrocellulose membrane, gold conjugate pad, sample pad, filter paper and absorbent pad were assembled to prepare the strips. The detection specificity and sensitivity of this strip were evaluated.

RESULTS

The strip test for detecting Vibrio vulnificus yielded results in 20 to 30 min. The detection sensitivity of the test was 2×10(6) CFU/ml. The strip showed no cross-reaction with other bacterial strains. The strips remained stable after preservation at 4 degrees celsius; for 4 months.

CONCLUSION

With a high specificity and sensitivity, this strip test is applicable in the detection of Vibrio vulnificus.

摘要

目的

建立一种快速检测创伤弧菌的试纸条检测方法。

方法

用创伤弧菌(ATCC27562)免疫家兔获得抗创伤弧菌多克隆抗体。通过柠檬酸钠还原氯金酸(HAuCl₄·4H₂O)制备胶体金,并与多克隆抗体结合作为检测试剂。将多克隆抗体和羊抗兔免疫球蛋白分别包被在同一张硝酸纤维素膜上,分别用于样品检测和质量控制。将硝酸纤维素膜、金标结合垫、样品垫、滤纸和吸水垫组装制备试纸条。对该试纸条的检测特异性和灵敏度进行评估。

结果

检测创伤弧菌的试纸条检测在20至30分钟内出结果。该检测的灵敏度为2×10⁶CFU/ml。该试纸条与其他菌株无交叉反应。试纸条在4℃保存4个月后仍保持稳定。

结论

该试纸条检测方法特异性高、灵敏度高,适用于创伤弧菌的检测。

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