Paoluzi O A, Rossi P, Montesano C, Bernardi S, Carnieri E, Marchione O P, Nardi F, Iacopini F, Pica R, Paoluzi P
Department of Microbiology, University of Tor Vergata, Italy.
Helicobacter. 2001 Jun;6(2):130-5. doi: 10.1046/j.1523-5378.2001.00019.x.
Infection with CagA-positive Helicobacter pylori may be diagnosed by detecting cagA gene by polymerase chain reaction assay (PCR) or serum antibodies against CagA by Western blot analysis. The aim of this study is to evaluate whether results of PCR and Western blot analysis are in agreement in CagA status assessment.
Thirty-six dyspeptic patients with unknown H. pylori status underwent upper gastrointestinal endoscopy to assess the presence of mucosal lesions and to collect six gastric biopsies (three from the antrum and three from the body) for evaluation of H. pylori infection (rapid urease test, histology and PCR for ureA gene) and gastritis. CagA status was assessed by PCR (cagA gene) on two biopsy specimens and by Western blot analysis of serum (CagA-antibodies) in each patient.
At endoscopy, nine patients showed normal mucosa, 15 a duodenal ulcer and 12 antral erosions. Twenty-eight patients were found to be H. pylori-positive and eight H. pylori-negative. Of the 28 H. pylori-positive patients, 17 were CagA-positive and five were CagA-negative by both methods, five were CagA-positive by Western blot analysis but not by PCR and one was CagA-positive by PCR but not by Western blot analysis. Of the eight H. pylori-negative patients, none was CagA-positive by PCR, while six were CagA-positive by Western blot analysis. Therefore, the two tests agreed in only 24 patients (67%). In those patients in whom the PCR and Western blot analysis were not in agreement, the histological features appear to suggest that the results of the Western blot analysis should be considered false positives or false negatives.
PCR and Western blot analysis failed to provide comparable data in many cases. Western blot analysis seems to be more likely to give misleading results than PCR. Thus, PCR seems to be the method of choice to assess CagA status.
通过聚合酶链反应检测法(PCR)检测cagA基因或通过蛋白质印迹分析检测抗CagA血清抗体,可诊断CagA阳性幽门螺杆菌感染。本研究旨在评估PCR和蛋白质印迹分析结果在CagA状态评估中是否一致。
36例幽门螺杆菌状态不明的消化不良患者接受了上消化道内镜检查,以评估黏膜病变情况,并采集6份胃活检组织(3份取自胃窦,3份取自胃体),用于评估幽门螺杆菌感染(快速尿素酶试验、组织学检查及ureA基因PCR检测)及胃炎情况。通过对两份活检标本进行PCR(检测cagA基因)及对每位患者血清进行蛋白质印迹分析(检测CagA抗体)来评估CagA状态。
在内镜检查中,9例患者黏膜正常,15例有十二指肠溃疡,12例有胃窦糜烂。28例患者幽门螺杆菌检测呈阳性,8例呈阴性。在28例幽门螺杆菌阳性患者中,两种方法均显示17例为CagA阳性,5例为CagA阴性,5例蛋白质印迹分析显示为CagA阳性但PCR检测为阴性,1例PCR检测显示为CagA阳性但蛋白质印迹分析为阴性。在8例幽门螺杆菌阴性患者中,PCR检测均未显示CagA阳性,而蛋白质印迹分析显示6例为CagA阳性。因此,两种检测方法仅在24例患者(67%)中结果一致。在PCR和蛋白质印迹分析结果不一致的患者中,组织学特征似乎表明蛋白质印迹分析结果应为假阳性或假阴性。
在许多情况下,PCR和蛋白质印迹分析未能提供可比数据。蛋白质印迹分析似乎比PCR更易产生误导性结果。因此,PCR似乎是评估CagA状态的首选方法。