Aoyama T, Kato T, Takeuchi Y, Kato K, Morokuma K, Hirai T
Department of Pediatrics, Kawasaki Municipal Hospital, Kawasaki-shi, Japan.
J Clin Microbiol. 1997 Jul;35(7):1859-61. doi: 10.1128/jcm.35.7.1859-1861.1997.
We developed a particle agglutination test (KPA) with poly(gamma-methyl L-glutamate) as the solid particle for measurement of pertussis toxin (PT) antibody. In this study, KPA was assessed as a means of serodiagnosing pertussis, and the results were compared with those of indirect enzyme-linked immunosorbent assay (indirect ELISA) and the microagglutination test. First, four serum samples were collected from each of 21 healthy children: before and 4 weeks after receiving three primary doses of acellular pertussis vaccines and before and 4 weeks after receiving a booster dose. In all 21 vaccinees, a significant rise in PT antibody titers was observed by KPA after each vaccination, and among all 84 serum samples collected, an excellent correlation was demonstrated between the values obtained by indirect ELISA and those obtained by KPA (r = 0.92). Second, paired serum samples were collected at intervals of approximately 2 weeks from 51 patients with culture-confirmed pertussis. A significant increase in titer (fourfold or more) was observed in 39 (76%) patients by KPA, 34 (67%) patients by indirect ELISA, and 23 (45%) patients by the microagglutination test. In acute- and convalescent-phase sera collected from 20 nonpertussis patients, there were no changes in titers by KPA. The KPA procedure was as simple as that of the microagglutination test, and the reaction time was only 2 h (or overnight). In this study, KPA was demonstrated to be a simple, speedy, sensitive, and specific serodiagnostic method for pertussis.
我们开发了一种以聚(γ-甲基-L-谷氨酸)为固相颗粒的颗粒凝集试验(KPA),用于检测百日咳毒素(PT)抗体。在本研究中,对KPA作为百日咳血清诊断方法进行了评估,并将结果与间接酶联免疫吸附测定(间接ELISA)和微量凝集试验的结果进行了比较。首先,从21名健康儿童中每人采集4份血清样本:在接种3剂无细胞百日咳疫苗的初次免疫前和免疫后4周,以及在接种加强剂量疫苗前和接种后4周。在所有21名接种疫苗者中,每次接种后通过KPA观察到PT抗体滴度显著升高,在采集的所有84份血清样本中,间接ELISA和KPA获得的值之间显示出极好的相关性(r = 0.92)。其次,从51例培养确诊的百日咳患者中每隔约2周采集配对血清样本。通过KPA观察到39例(76%)患者滴度显著升高(四倍或更高),通过间接ELISA观察到34例(67%)患者滴度显著升高,通过微量凝集试验观察到23例(45%)患者滴度显著升高。在从20例非百日咳患者采集的急性期和恢复期血清中,KPA检测的滴度没有变化。KPA操作与微量凝集试验一样简单,反应时间仅为2小时(或过夜)。在本研究中,KPA被证明是一种用于百日咳的简单、快速、灵敏且特异的血清诊断方法。