Nested primers improve sensitivity in the detection of Helicobacter pylori by the polymerase chain reaction

被引:30
作者
Bamford, KB
Lutton, DA
O'Loughlin, B
Coulter, WA
Collins, JSA
机构
[1] Queens Univ, Dept Microbiol & Immunobiol, Belfast BT12 6BN, Antrim, North Ireland
[2] Queens Univ, Sch Dent, Belfast BT12 6BN, Antrim, North Ireland
[3] Royal Victoria Hosp, Belfast BT12 6BA, Antrim, North Ireland
关键词
D O I
10.1016/S0163-4453(98)93450-8
中图分类号
R51 [传染病];
学科分类号
100401 ;
摘要
To investigate potential routes of spread of infection by the polymerase chain reaction (PCR) it is important that the technique is effective in the types of specimen to be investigated. To establish the limits of detection of Helicobacter pylori by PCR in clinical material from the gastric mucosa, faeces, dental plaque and oral rinses, samples were seeded with known numbers of bacteria. DNA extraction was followed by amplification with primers from the urease C gene. Nested primers were used to amplify the PCR product which was detected using a digoxigenin-labelled probe. Faeces or plaque inhibited the single reaction 10(2)-10(6) fold, A second amplification using nested primers and probing increased the sensitivity to a level similar to that obtained with pure culture. This method is potentially useful with less likelihood of false negative results when trying to detect H. pylori by PCR in highly contaminated, clinical material.
引用
收藏
页码:105 / 110
页数:6
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