Durkin M M, Bartlett M S, Queener S F, Shaw M M, Lee C H, Smith J W
Department of Pathology, Indiana University School of Medicine, Indianapolis 46202.
J Clin Microbiol. 1992 Dec;30(12):3258-62. doi: 10.1128/jcm.30.12.3258-3262.1992.
An enzyme-linked immunosorbent assay (ELISA) to quantitate Pneumocystis carinii organisms from culture supernatant and rat lung has been developed. A polyclonal antibody specific to P. carinii was produced in Sprague-Dawley rats by allowing P. carinii-infected animals to recover from infection. This antibody reacted strongly to P. carinii proteins of 50 to 55 kDa and weakly to those of 33 and 116 kDa. The ELISA used this convalescent-phase antibody to quantitate the number of P. carinii organisms in lung homogenates of infected rats and supernatants from infected tissue cultures which were used to screen drugs for P. carinii. The results of the ELISA were compared with those of direct microscopic counting of organisms, and the two methods were highly correlated (r > 0.9). Thus, the ELISA can be used as an alternative method for the quantitation of P. carinii organisms, and it is superior to the conventional microscopic method because it is easier to perform and less labor-intensive.
已开发出一种酶联免疫吸附测定法(ELISA),用于定量培养上清液和大鼠肺中的卡氏肺孢子虫。通过让感染卡氏肺孢子虫的动物从感染中恢复,在斯普拉格-道利大鼠中产生了一种对卡氏肺孢子虫特异的多克隆抗体。该抗体与50至55 kDa的卡氏肺孢子虫蛋白强烈反应,与33和116 kDa的蛋白反应较弱。ELISA使用这种恢复期抗体来定量感染大鼠肺匀浆和感染组织培养上清液中卡氏肺孢子虫的数量,这些上清液用于筛选治疗卡氏肺孢子虫的药物。将ELISA的结果与直接显微镜计数生物体的结果进行比较,两种方法高度相关(r>0.9)。因此,ELISA可作为定量卡氏肺孢子虫生物体的替代方法,并且它优于传统的显微镜方法,因为它更易于操作且劳动强度较小。