Masayoshi T, Hirai Y, Kanemasa Y
Department of Microbiology, Okayama University Medical School, Japan.
Microbiol Immunol. 1992;36(2):149-60. doi: 10.1111/j.1348-0421.1992.tb01652.x.
We prepared polyclonal antibody specific to Mycoplasma pneumoniae. Using this antibody, we developed a latex agglutination test (LAT) for detecting the organism in respiratory exudates as rapid diagnosis of M. pneumoniae infection. Further, LAT was compared with DNA-probe test (DP) which was the only commercially available test for the rapid detection of the organism. In LAT, both M. pneumoniae and M. genitalium give positive agglutination, but the titer of M. genitalium was significantly lower than that of M. pneumoniae. The detection limit of LAT was 2 x 10(5) CFU/ml and that of DP was 5 x 10(4) CFU/ml in vitro. It was considered that target molecules in LAT were accumulated in the pharyngeal portion of the patients, because of their long half-life at 37 C. However, ribosomal RNA which was target molecule in DP was destroyed at 37 C much sooner, and the accumulation could not be expected. Actually, positive rate in LAT was higher than that in DP among clinical specimens in which M. pneumoniae was detected by culture method. The procedure of LAT is much easier and more rapid than that of DP in which radioactive isotope is required. LAT could be the choice of test for rapid diagnosis of M. pneumoniae infection.
我们制备了针对肺炎支原体的多克隆抗体。利用该抗体,我们开发了一种乳胶凝集试验(LAT),用于检测呼吸道分泌物中的该病原体,以快速诊断肺炎支原体感染。此外,将LAT与DNA探针试验(DP)进行了比较,DP是当时唯一可用于快速检测该病原体的商业可用试验。在LAT中,肺炎支原体和生殖支原体均呈阳性凝集,但生殖支原体的滴度明显低于肺炎支原体。LAT的体外检测限为2×10⁵CFU/ml,DP的体外检测限为5×10⁴CFU/ml。据认为,LAT中的靶分子在患者咽部蓄积,因为它们在37℃时有较长的半衰期。然而,DP中的靶分子核糖体RNA在37℃时更快地被破坏,因此无法预期会有蓄积。实际上,在通过培养方法检测到肺炎支原体的临床标本中,LAT的阳性率高于DP。LAT的操作程序比需要放射性同位素的DP更容易、更快速。LAT可作为快速诊断肺炎支原体感染的检测方法。