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通过源自新克隆DNA探针的巢式聚合酶链反应检测法鉴定唾液中的幽门螺杆菌。

Identification of H. pylori in saliva by a nested PCR assay derived from a newly cloned DNA probe.

作者信息

Jiang C, Li C, Ha T, Ferguson D A, Chi D S, Laffan J J, Thomas E

机构信息

Department of Internal Medicine, James H. Quillen College of Medicine, East Tennessee State University, and Veterans Affairs Medical Center, Johnson City 37684, USA.

出版信息

Dig Dis Sci. 1998 Jun;43(6):1211-8. doi: 10.1023/a:1018847522200.

Abstract

A novel probe was developed from genomic DNA of Helicobacter pylori ATCC type strain 43629. It hybridized with all 73 H. pylori clinical isolates tested but not with any of 183 non-H. pylori DNAs in dot blot hybridization. Typing tests revealed 41 different HaeIII-digestion patterns from 57 H. pylori strains tested. Based on the sequence of the probe, a nested PCR was developed that detected as little as 2 fg of H. pylori DNA or approximately equivalent to one cell. No PCR products were amplified from any of 21 non-H. pylori strains tested. Using this nested PCR, H. pylori DNA was detected in 33 of 45 (73%) saliva samples collected from patients with gastric H. pylori infection. These data suggest that the probe is useful for typing H. pylori and that the nested PCR is a valuable tool for detecting H. pylori DNA in saliva.

摘要

一种新型探针是从幽门螺杆菌ATCC标准菌株43629的基因组DNA中开发出来的。在斑点印迹杂交中,它与所检测的全部73株幽门螺杆菌临床分离株杂交,但不与183种非幽门螺杆菌DNA中的任何一种杂交。分型试验显示,在检测的57株幽门螺杆菌菌株中出现了41种不同的HaeIII酶切模式。根据探针序列,开发了一种巢式PCR,该方法可检测低至2 fg的幽门螺杆菌DNA,约相当于一个细胞。在检测的21种非幽门螺杆菌菌株中均未扩增出PCR产物。使用这种巢式PCR,在从幽门螺杆菌感染患者采集的45份唾液样本中的33份(73%)中检测到了幽门螺杆菌DNA。这些数据表明,该探针可用于幽门螺杆菌分型,巢式PCR是检测唾液中幽门螺杆菌DNA的有价值工具。

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