Danielová V, Holubová J, Sobotková J
Státní zdravotní ústav, Praha.
Epidemiol Mikrobiol Imunol. 1996 May;45(2):59-67.
The authors tested the suitability of 10 cell lines for assessment of the virulence of strains of Yersinia enterocolitica by the cytotoxicity test. Line LLC-MK2, HEp-2, HeLa, L-132 and L929 are most suitable. Lines LEP, 3T3, MDCK and McCoy are usable, line BHK-21 is unsuitable. On the same cell lines also the invasiveness of the virulent and non-virulent strains of Y. enterocolitica was assessed. For detection of the internalization of bacteria a modified method of double fluorescent staining was used which is simpler and more rapid. The invasiveness, similarly as the cytotoxicity of the virulent strain was proved on all tested cell lines, contrary to the non-virulent strain which was neither cytotoxic nor invasive. Using strains of Y. enterocolitica, isolated from faeces of patients with diarrhoea of infectious etiology or another bowel disease and from water, the cytotoxicity method was compared with recommended biochemical virulence tests used in this country: bond with Congo red, crystal violet, dependence on Ca2+ and pyrazinamidase activity. Strains of serotype 0:5, 0:6.31 and 0:7.8 biotype 1 gave in all tests negative results, strains of serotype 0:3 biotype 4 differed. The highest number of positive strains was detected by the cytotoxicity test. The positivity of strains of this serotype was assessed by the invasiveness test in vitro. The cytotoxicity test was thus proved to be a very effective, rapid and easy test of detection of the virulence of Y. enterocolitica strains.
作者通过细胞毒性试验测试了10种细胞系对小肠结肠炎耶尔森氏菌菌株毒力评估的适用性。LLC-MK2、HEp-2、HeLa、L-132和L929细胞系最为合适。LEP、3T3、MDCK和McCoy细胞系可用,BHK-21细胞系不合适。在相同的细胞系上,还评估了小肠结肠炎耶尔森氏菌有毒和无毒菌株的侵袭性。为检测细菌的内化,使用了一种改良的双荧光染色方法,该方法更简单、更快速。与无毒菌株不同,无毒菌株既无细胞毒性也无侵袭性,在所有测试的细胞系上都证明了有毒菌株的侵袭性与细胞毒性相似。使用从感染性病因腹泻或其他肠道疾病患者粪便以及水中分离出的小肠结肠炎耶尔森氏菌菌株,将细胞毒性方法与该国推荐的生化毒力试验进行了比较:与刚果红、结晶紫结合,对Ca2+的依赖性以及吡嗪酰胺酶活性。0:5、0:6.31和0:7.8生物型1血清型的菌株在所有试验中均给出阴性结果,0:3生物型4血清型的菌株则不同。细胞毒性试验检测到的阳性菌株数量最多。通过体外侵袭试验评估了该血清型菌株的阳性率。因此,细胞毒性试验被证明是检测小肠结肠炎耶尔森氏菌菌株毒力的一种非常有效、快速且简便的试验。