一种用于检测溶血巴斯德氏菌白细胞毒素的比色微孔板测定法。

A colourimetric, microplate assay for the leucotoxin of Pasteurella haemolytica.

作者信息

Craig F F, Dalgleish R, Sutherland A D, Parton R, Coote J G, Gibbs H A, Freer J H

机构信息

Dept. of Microbiology, University of Glasgow, Great Britain.

出版信息

Vet Microbiol. 1990 May;22(4):309-17. doi: 10.1016/0378-1135(90)90017-p.

DOI:10.1016/0378-1135(90)90017-p

PMID:2363244

Abstract

Culture supernates of Pasteurella haemolytica, which contain leucotoxin, inhibited the reduction of nitroblue tetrazolium (NBT) by bovine and ovine but not rabbit leucocytes in response to phorbol 12-myristate 13-acetate (PMA). Culture supernates of P. multocida, which contain no leucotoxin, had no inhibitory effect on the response of leucocytes from any species. The inhibition of NBT reduction was assessed visually or spectrophotometrically in the wells of microplates and used as a simple assay for leucotoxin. It was as sensitive as the trypan blue dye-exclusion method and did not require the use of radioisotopes. In addition, sera from P. haemolytica-infected calves inhibited leucotoxin activity in the microplate assay. Thus, inhibition of NBT reduction after stimulation of ruminant leucocytes with PMA can be used as a simple, specific assay for leucotoxin and for anti-leucotoxin antibodies.

摘要

含有白细胞毒素的溶血巴斯德菌培养上清液，可抑制佛波醇12 -肉豆蔻酸酯13 -乙酸酯（PMA）刺激后牛和羊白细胞对硝基蓝四氮唑（NBT）的还原反应，但对兔白细胞无此作用。不含白细胞毒素的多杀巴斯德菌培养上清液，对任何物种的白细胞反应均无抑制作用。通过在微孔板孔中进行目视或分光光度法评估NBT还原的抑制情况，并将其用作白细胞毒素的简单检测方法。该方法与台盼蓝染料排除法一样灵敏，且无需使用放射性同位素。此外，来自感染溶血巴斯德菌的犊牛的血清在微孔板检测中可抑制白细胞毒素活性。因此，用PMA刺激反刍动物白细胞后对NBT还原的抑制作用，可作为白细胞毒素和抗白细胞毒素抗体的一种简单、特异性检测方法。