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一种用于鉴定牙菌斑和胃活检标本中幽门螺杆菌的快速聚合酶链反应方法的开发。

Development of a rapid PCR method for identification of Helicobacter pylori in dental plaque and gastric biopsy specimens.

作者信息

Wahlfors J, Meurman J H, Toskala J, Korhonen A, Alakuijala P, Janatuinen E, Kärkkkäinen U M, Nuutinen P, Jänne J

机构信息

Department of Biochemistry and Biotechnology, A.I. Virtanen Institute, Kuopio, Finland.

出版信息

Eur J Clin Microbiol Infect Dis. 1995 Sep;14(9):780-6. doi: 10.1007/BF01690993.

Abstract

A rapid and simple polymerase chain reaction (PCR) method was developed to detect Helicobacter pylori in gastric biopsy specimens and dental plaque samples. The primers were targeted to the 16S rRNA sequence of Helicobacter pylori strain ATCC 43504. The system was found to have a theoretical detection level of 0.5 to 5 Helicobacter pylori cells in a 5 microliters sample of dental plaque. In the absence of plaque, the detection level was even better: theoretically, 0.05 to 0.5 Helicobacter pylori cells were detected in water suspension. However, this appeared to be due to the presence of free bacterial DNA in the culture used for the sensitivity determination. Thus, the actual sensitivity of the system was found to be fewer than five Helicobacter pylori cells, irrespective of the type of sample used. The method was then used to analyse 29 dental plaque and gastric biopsy specimens collected from patients with a history of recurrent peptic ulcer disease. Fourteen stomach specimens were positive for Helicobacter pylori when tested with the PCR method, while the respective figures with culture, histological examination and the urease test were 11, 12 and 9. No positive dental plaque samples were observed.

摘要

开发了一种快速简便的聚合酶链反应(PCR)方法,用于检测胃活检标本和牙菌斑样本中的幽门螺杆菌。引物靶向幽门螺杆菌菌株ATCC 43504的16S rRNA序列。该系统在5微升牙菌斑样本中的理论检测水平为0.5至5个幽门螺杆菌细胞。在没有牙菌斑的情况下,检测水平甚至更好:理论上,在水悬浮液中可检测到0.05至0.5个幽门螺杆菌细胞。然而,这似乎是由于用于灵敏度测定的培养物中存在游离细菌DNA。因此,无论使用何种类型的样本,该系统的实际灵敏度均低于5个幽门螺杆菌细胞。然后使用该方法分析了从有复发性消化性溃疡病史的患者中收集的29个牙菌斑和胃活检标本。用PCR方法检测时,14个胃标本的幽门螺杆菌呈阳性,而培养、组织学检查和尿素酶试验的相应数字分别为11、12和9。未观察到阳性牙菌斑样本。

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