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组织和尿液自溶产物对波摩那群问号钩端螺旋体的活力、形态及抗原性的影响

Effects of products of autolysis of tissue and urine on the viability, morphology, and antigenicity of Leptospira interrogans serovar pomona.

作者信息

Fairbrother J M

出版信息

J Clin Microbiol. 1985 Feb;21(2):189-94. doi: 10.1128/jcm.21.2.189-194.1985.

Abstract

The effect of the products of autolysis on Leptospira interrogans serovar pomona was investigated in bovine kidney tissues and urine inoculated with this organism. No viable leptospires were found at 24 h or subsequently after inoculation of kidney tissues or urine. Leptospires and their soluble antigens were rapidly destroyed in tissues stored at 20 degrees C and could not be detected by the fluorescent-antibody test and the enzyme-linked immunosorbent assay. On the other hand, leptospires were detectable by dark-field microscopy and fluorescent-antibody test 38 days after addition to urine. The rate of destruction proceeded less rapidly in tissues stored at 4 degrees C. Under the conditions of this study, soluble leptospiral antigen levels decreased less rapidly than did the number of detectable leptospires. Similarly, breakdown of leptospires, demonstrated by dark-field microscopy and fluorescent-antibody test, progressed during the first 5 days after inoculation of urine, when levels of soluble leptospiral antigen remained constant. Although the number of intact leptospires was markedly reduced after freezing of tissues at -20 degrees C, soluble leptospiral antigen levels remained unaltered. Neither intact leptospires nor their soluble antigens were detected after preservation of tissues in Formalin. However, formolization of urine delayed the destruction of leptospires and increased the levels of soluble leptospiral antigen during at least the first 5 days after inoculation. These results indicate that assays such as the enzyme-linked immunosorbent assay should be useful for the detection of soluble leptospiral antigens in urine and in autolyzing tissues such as those taken from a bovine fetus aborted as a result of leptospiral infection.

摘要

在接种问号钩端螺旋体波摩那群的牛肾组织和尿液中,研究了自溶产物对该菌的影响。接种肾组织或尿液后24小时及之后未发现存活的钩端螺旋体。钩端螺旋体及其可溶性抗原在20℃保存的组织中迅速被破坏,无法通过荧光抗体试验和酶联免疫吸附测定检测到。另一方面,加入尿液38天后,通过暗视野显微镜检查和荧光抗体试验可检测到钩端螺旋体。在4℃保存的组织中,破坏速度较慢。在本研究条件下,可溶性钩端螺旋体抗原水平下降速度比可检测到的钩端螺旋体数量下降速度慢。同样,接种尿液后的前5天,通过暗视野显微镜检查和荧光抗体试验证明钩端螺旋体的分解过程中,可溶性钩端螺旋体抗原水平保持恒定。虽然在-20℃冷冻组织后完整钩端螺旋体的数量明显减少,但可溶性钩端螺旋体抗原水平保持不变。用福尔马林保存组织后,既未检测到完整的钩端螺旋体,也未检测到其可溶性抗原。然而,尿液的甲醛固定至少在接种后的前5天延迟了钩端螺旋体的破坏,并增加了可溶性钩端螺旋体抗原的水平。这些结果表明,诸如酶联免疫吸附测定等检测方法对于检测尿液和自溶组织(如因钩端螺旋体感染而流产的牛胎儿的组织)中的可溶性钩端螺旋体抗原应该是有用的。

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1
Ultra-rapid fluorescent labelling of proteins.蛋白质的超快速荧光标记
Nature. 1962 Jan 13;193:167-8. doi: 10.1038/193167b0.
10
Leptospirosis.钩端螺旋体病
Vet Rec. 1970 Apr 25;86(17):484-96. doi: 10.1136/vr.86.17.484.

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