Rokosz-Chudziak Natalia, Rastawicki Waldemar
Med Dosw Mikrobiol. 2014;66(3-4):195-207.
Campylobacterjejuni is has been found to be the leading cause of bacterial gastroenteritis worldwide. Clinical manifestation on enterocolitis caused by C. jejuni are diarrhea, fever, abdominal pain and in some patients, fecal blood. After C. jejuni infection, squeals may occur such as reactive arthritis. The aim of the study was to investigate the frequency of antibodies to the recombinant protein P39 sticks C. jejuni in patients with gastrointestinal disorders and reactive arthritis in Poland.
Serum samples collected from 46 patients with bacteriology confir- med infection caused by Campylobacter jejuni, 472 sera from patients with gastrointestinal disorders, 97 serum samples obtained from patients with reactive arthritis and 84 sera from healthy adults and children. Sera were screened for anti-P39 C. jejuni recombinant protein IgA, IgG andIgM antibodies by using the home-made ELISA. Protein P39 C. jejuni was expressing in E. coli BL21 (DE3) using the pET-30 Ek/LIC expression vector. Purification was accomplished by immobilized metal (Ni2+) affinity column chromatography (His Bind Resign, Novagen).
SDS-PAGE and Coomassie brilliant blue staining confirmed a high purity of the recombinant P39 protein preparation with an expected molecular mass of 39 kDa. The results of ELISA with the P39 recombinant protein revealed that IgA antibodies in diagnostically significant level (x + 2SD) were found in 18.8%, IgM in 14.8% and IgG in 7.8% of sera obtained from patients with of gastrointestinal disorders. On the other hand, antibodies to recombinant P39 protein in sera obtained from patients with reactive arthritis were found in more than twice the percentage than in patients with gastrointestinal disorders (IgA in 34.0%, IgG in 26.8% and IgM in 19.6%).
In conclusion, based on the data obtained, C. jejuni may be important factor in triggering the gastrointestinal disorder and reactive arthritis in humans in Poland.
空肠弯曲菌已被发现是全球细菌性肠胃炎的主要病因。空肠弯曲菌引起的小肠结肠炎的临床表现为腹泻、发热、腹痛,部分患者还会出现便血。空肠弯曲菌感染后,可能会出现诸如反应性关节炎等并发症。本研究的目的是调查波兰胃肠道疾病患者和反应性关节炎患者中针对空肠弯曲菌重组蛋白P39的抗体频率。
收集46例经细菌学确诊为空肠弯曲菌感染患者的血清样本、472例胃肠道疾病患者的血清、97例反应性关节炎患者的血清样本以及84例健康成人和儿童的血清。采用自制酶联免疫吸附测定法(ELISA)检测血清中抗空肠弯曲菌重组蛋白P39的IgA、IgG和IgM抗体。使用pET - 30 Ek/LIC表达载体在大肠杆菌BL21(DE3)中表达空肠弯曲菌蛋白P39。通过固定金属(Ni2 +)亲和柱色谱法(His Bind Resign,诺华公司)完成纯化。
十二烷基硫酸钠 - 聚丙烯酰胺凝胶电泳(SDS - PAGE)和考马斯亮蓝染色证实重组P39蛋白制剂具有高纯度,预期分子量为39 kDa。用P39重组蛋白进行ELISA的结果显示,在胃肠道疾病患者的血清中,18.8%的血清中IgA抗体达到诊断显著水平(x + 2SD),14.8%的血清中IgM抗体达到该水平,7.8%的血清中IgG抗体达到该水平。另一方面,反应性关节炎患者血清中针对重组P39蛋白的抗体比例是胃肠道疾病患者的两倍多(IgA为34.0%,IgG为26.8%,IgM为19.6%)。
总之,根据所获得的数据,空肠弯曲菌可能是引发波兰人类胃肠道疾病和反应性关节炎的重要因素。