Uesaka Y, Otsuka Y, Kashida M, Oku Y, Horigome K, Nair G B, Pal S C, Yamasaki S, Takeda Y
Institute for Diagnostic Reagents, Nissui Pharmaceutical Co., Ibaraki, Japan.
Microbiol Immunol. 1992;36(1):43-53. doi: 10.1111/j.1348-0421.1992.tb01641.x.
A bead-enzyme linked immunosorbent assay (bead-ELISA) for detection and quantification of cholera toxin (CT) in broth cultures of Vibrio cholerae O1 has been developed. Under optimal buffer and pH conditions the bead-ELISA could consistently detect 40 pg/ml of CT. None of the ingredients of commonly used media for in vitro culture of V. cholerae O1 hindered the performance of the bead-ELISA. Evaluation of the sensitivity and specificity of the bead-ELISA against the commonly used reversed passive latex agglutination (RPLA) test for detection of CT was performed using a collection of 239 strains of V. cholerae O1 (including both biotypes and serotypes) which were examined by a gene probe encoding for the A1 subunit of CT. Although both the assays were highly specific, the bead-ELISA was more sensitive than the RPLA. Quantification of CT by the bead-ELISA revealed that the concentration of CT produced by the strains of V. cholerae O1 which were negative by the RPLA was lower than 1 ng/ml and therefore below the minimum detection ability of the RPLA. The bead-ELISA is a simple, specific and highly sensitive assay for routine detection of CT and is recommended for routine use in clinical microbiology laboratories.
已开发出一种用于检测和定量霍乱弧菌O1肉汤培养物中霍乱毒素（CT）的磁珠酶联免疫吸附测定法（磁珠ELISA）。在最佳缓冲液和pH条件下，磁珠ELISA能够始终如一地检测到40 pg/ml的CT。霍乱弧菌O1体外培养常用培养基的成分均未干扰磁珠ELISA的性能。使用一组239株霍乱弧菌O1（包括两种生物型和血清型）对磁珠ELISA相对于常用的反向被动乳胶凝集试验（RPLA）检测CT的敏感性和特异性进行了评估，这些菌株通过编码CT A1亚基的基因探针进行检测。尽管两种检测方法都具有高度特异性，但磁珠ELISA比RPLA更敏感。通过磁珠ELISA对CT进行定量分析发现，RPLA检测呈阴性的霍乱弧菌O1菌株产生的CT浓度低于1 ng/ml，因此低于RPLA的最低检测能力。磁珠ELISA是一种用于常规检测CT的简单、特异且高度灵敏的检测方法，推荐在临床微生物实验室中常规使用。
