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CLC number: R722 13
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Received: 1999-05-05
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DETECTION OF BACTERIAL DNA BY PCR AND REVERSE HYBRIDIZATION IN THE 16s rRNA GENE
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SHANG Shi-qiang, YU Xi-lin, HONG Wen-lan, YU Hui-min, SUN Mei-yue. DETECTION OF BACTERIAL DNA BY PCR AND REVERSE HYBRIDIZATION IN THE 16s rRNA GENE[J]. Journal of Zhejiang University Science A, 2000, 1(2): 222-226.
@article{title="DETECTION OF BACTERIAL DNA BY PCR AND REVERSE HYBRIDIZATION IN THE 16s rRNA GENE",
author="SHANG Shi-qiang, YU Xi-lin, HONG Wen-lan, YU Hui-min, SUN Mei-yue",
journal="Journal of Zhejiang University Science A",
volume="1",
number="2",
pages="222-226",
year="2000",
publisher="Zhejiang University Press & Springer",
doi="10.1631/jzus.2000.0222"
}
%0 Journal Article
%T DETECTION OF BACTERIAL DNA BY PCR AND REVERSE HYBRIDIZATION IN THE 16s rRNA GENE
%A SHANG Shi-qiang
%A YU Xi-lin
%A HONG Wen-lan
%A YU Hui-min
%A SUN Mei-yue
%J Journal of Zhejiang University SCIENCE A
%V 1
%N 2
%P 222-226
%@ 1869-1951
%D 2000
%I Zhejiang University Press & Springer
%DOI 10.1631/jzus.2000.0222
TY - JOUR
T1 - DETECTION OF BACTERIAL DNA BY PCR AND REVERSE HYBRIDIZATION IN THE 16s rRNA GENE
A1 - SHANG Shi-qiang
A1 - YU Xi-lin
A1 - HONG Wen-lan
A1 - YU Hui-min
A1 - SUN Mei-yue
J0 - Journal of Zhejiang University Science A
VL - 1
IS - 2
SP - 222
EP - 226
%@ 1869-1951
Y1 - 2000
PB - Zhejiang University Press & Springer
ER -
DOI - 10.1631/jzus.2000.0222
Abstract: The clinical diagnosis of sepsis is difficult, particularly in neonates. To devise a rapid and reliable method for identifying bacteria in blood and cerebrospinal fluid (CSF), we developed a pair of primers according to the gene encoding 16 s rRNA, found in all bacteria. DNA fragments from different bacterial species and from clinical samples were detected with polymerase chain reaction (PCR), and with reverse hybridization using a universal bacterial probe, a gram-positive probe and a gram-negative probe. Our results showed that a 371 bp DNA fragment was amplified from 20 different bacterial species. No signal was observed when human DNA and viruses were used as templates. The sensitivity could be improved to 10-12 g. All 26 culture-positive clinical samples (22 blood samples and 4 CSF samples), were positive with PCR. The gram-negative and gram-positive probe hybridized to clinical samples and to known bacterial controls, as predicted by Gram
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