| 宋敏,李进,马维芳,徐湘民,张基增.应用巢式聚合酶链反应在唾液中检出幽门螺杆菌[J].Chinese journal of Epidemiology,1993,14(4):237-240 |
| 应用巢式聚合酶链反应在唾液中检出幽门螺杆菌 |
| Detection of Helicobacter pylari in Human Saliva by Using Nested Polymerase Chain Reaction |
| Received:February 02, 1993 Revised:March 05, 1993 |
| DOI： |
| KeyWord: 幽门螺杆菌 巢式聚合酶链反应 唾液 |
| English Key Word: Helicobacter pylori Nested PCR Saliva |
| FundProject: |
| Author Name | Affiliation | | Song Min | Dept. of Histology and Embryology, First Medical University of PLA, Guangzhou 510515 | | 李进 | Dept. of Histology and Embryology, First Medical University of PLA, Guangzhou 510515 | | 马维芳 | Dept. of Histology and Embryology, First Medical University of PLA, Guangzhou 510515 | | 徐湘民 | Dept. of Histology and Embryology, First Medical University of PLA, Guangzhou 510515 | | 张基增 | Dept. of Histology and Embryology, First Medical University of PLA, Guangzhou 510515 |
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| Abstract: |
| 用互补于幽门螺杆菌(HP)尿素酶A基因的两对引物行巢式聚合酶链反应(N-PCR),检测1株HP标准菌株、20株HP临床分离株均阳性,而12种肠道菌均阴性,特异性100%。该法敏感性好可检测0.1fg细菌DNA.57例因上消化道症状行胃镜检查者,取粘膜分别做细菌培养、尿素酶试验、组织学检查和N-PCR检测,其中27例在行胃镜前收集其唾液标本做N-PCR。19例胃粘膜HP阳性者中有11例唾液中检出HP,而8例胃粘膜HP阴性者中有1例唾液N-PCR阳性。作者认为口腔中确实存在HP,该菌可能通过口—口途径传播。 |
| English Abstract: |
| Epidemiologic studies suggest person to person spread of Helicobacter pylori (H. pylori), but the exact mechanism is unknown. Spread through oral secretions has been suggested, however it has proved very difficult to grow the organism from areas outside the stomach. A nested polymerase chain reaction (N-PCR) for the specific detection of H. pylori was developed with two primer pairs (nested primers) derived from the urease gene A of H. pylori. The N-PCR could detect all 21 H. pylori strains, including 20 isolated strains and 1 reference strain NCTC 14123, but could not detect other bacterial species, showing the N-PCR assay to be 100% specific. Tenfold serial dilution experimemts revealed the detection of as little as 0.1fg DNA by N-PCR. To evaluate the PCR assay for clinical samples, gastric biopsy and aspirate specimens were tested by N-PCR, and the results were compared with those of culture, urease test and histologic examination (reference standard, RS). Among 57 biopsy specimens, H. pylori sequence was detected by PCR in 39 of 39 (100%) positive tissues and in none of 18 negative tissues. H. pylori was detected in saliva of 11 out of 19 cases in which H. pylori was positive in gastric mucosa by PCR. Whereas, PCR was positive in saliva of only one out of 8 cases in which H. pylori was negative in gastric mucosa. Six gastric aspirate specimens were positive by N-PCR. PCR is an accurate and sensitive method that can detect the presence of H. pylori without the need of culture. |
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