Catalano M, Rosetti F A, Ochoa L B
Departamento de Microbiología, Facultad de Medicina, Universidad de Buenos Aires, Argentina.
Rev Argent Microbiol. 1990 Jul-Sep;22(3):115-22.
Sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) applied to outer membrane protein (OMP), extracted by a micromethod, was employed to subtype H. influenza b type I. A total of 37 H. influenzae b strains were isolated from children under 4 years of age, either with lower acute respiratory infection (LARI), or asymptomatic carriers matched according sex, socioeconomic level and seasonality. Twenty seven out of the 37 H. influenzae b strains belonged to biotype I. On the basis of OMP profiles, these 27 were classified into 8 subtypes (Figs. 2, 3, 4 and 5). The probability of two randomly chosen isolates having different OMP profiles was 0.733. The subtype termed "a" showed the greatest relative frequency and was detected both in invasive strains and in those isolated from throat samples of LARI cases and healthy children. The use of 14% SDS-PAGE allowed de detection either of a 51kD or a 49kD, as well as 25-40kD proteins, in a single run (Fig. 1). Most subtype profiles showed the 51 kD protein. Growth conditions and extraction of OMPs by our modified micromethod provide a single and inexpensive procedure within the means of the average clinical laboratory. Besides, this test is much less time-consuming than classical assays. Jointly, biotyping , serotyping and OMP profile determination, proved a useful epidemiological tool to survey H. influenzae b infection.
采用微量方法提取外膜蛋白(OMP)后,通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)对I型b型流感嗜血杆菌进行亚型分析。从4岁以下患有下呼吸道急性感染(LARI)的儿童或按性别、社会经济水平和季节匹配的无症状携带者中,共分离出37株b型流感嗜血杆菌。37株b型流感嗜血杆菌中,有27株属于生物I型。根据OMP图谱,将这27株分为8个亚型(图2、3、4和5)。随机选择的两个分离株具有不同OMP图谱的概率为0.733。称为“a”的亚型显示出最高的相对频率,在侵袭性菌株以及从LARI病例和健康儿童的咽喉样本中分离出的菌株中均有检测到。使用14%的SDS-PAGE可以在一次电泳中检测到51kD或49kD以及25 - 40kD的蛋白质(图1)。大多数亚型图谱显示有51kD的蛋白质。我们改良的微量方法所采用的生长条件和OMP提取方法,为普通临床实验室提供了一种简单且成本低廉的程序。此外,该检测比传统检测耗时少得多。生物分型、血清分型和OMP图谱测定共同证明是一种用于调查b型流感嗜血杆菌感染的有用的流行病学工具。