DETECTION OF LOW NUMBERS OF BACTERIAL-CELLS IN SOILS AND SEDIMENTS BY POLYMERASE CHAIN-REACTION

被引:280
作者
TSAI, YL [1 ]
OLSON, BH [1 ]
机构
[1] UNIV CALIF IRVINE,PROGRAM SOCIAL ECOL,IRVINE,CA 92717
关键词
D O I
10.1128/AEM.58.2.754-757.1992
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
Polymerase chain reaction was used to amplify the low copy number of two 16S ribosomal gene fragments from soil and sediment extracts. Total DNA for polymerase chain reaction was extracted from 1 g of seeded or unseeded samples by a rapid freeze-and-thaw method. Amplified DNA fragments can be detected in DNA fractions isolated from seeded soil containing less than 3 Escherichia coli cells and from seeded sediments containing less than 10 cells. This research demonstrated that coupling polymerase chain reaction to direct DNA extraction improves sensitivity by 1 and 2 orders of magnitude for sediments and soils, respectively. This technique could become a powerful tool for genetic ecology studies.
引用
收藏
页码:754 / 757
页数:4
相关论文
共 19 条
[1]   DETECTION OF VIABLE LEGIONELLA-PNEUMOPHILA IN WATER BY POLYMERASE CHAIN-REACTION AND GENE PROBE METHODS [J].
BEJ, AK ;
MAHBUBANI, MH ;
ATLAS, RM .
APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 1991, 57 (02) :597-600
[2]   DETECTION OF COLIFORM BACTERIA IN WATER BY POLYMERASE CHAIN-REACTION AND GENE PROBES [J].
BEJ, AK ;
STEFFAN, RJ ;
DICESARE, J ;
HAFF, L ;
ATLAS, RM .
APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 1990, 56 (02) :307-314
[3]   GENE ORGANIZATION AND PRIMARY STRUCTURE OF A RIBOSOMAL-RNA OPERON FROM ESCHERICHIA-COLI [J].
BROSIUS, J ;
DULL, TJ ;
SLEETER, DD ;
NOLLER, HF .
JOURNAL OF MOLECULAR BIOLOGY, 1981, 148 (02) :107-127
[4]   MAXIMIZING SENSITIVITY AND SPECIFICITY OF PCR BY PREAMPLIFICATION HEATING [J].
DAQUILA, RT ;
BECHTEL, LJ ;
VIDELER, JA ;
ERON, JJ ;
GORCZYCA, P ;
KAPLAN, JC .
NUCLEIC ACIDS RESEARCH, 1991, 19 (13) :3749-3749
[5]  
DAVIS LG, 1986, BASIC METHODS MOL BI, P147
[6]  
DELEON R, 1991, P WAT QUAL TECHN C S, V18, P833
[7]   DETECTION OF CYTOMEGALO-VIRUS IN URINE FROM NEWBORNS BY USING POLYMERASE CHAIN-REACTION DNA AMPLIFICATION [J].
DEMMLER, GJ ;
BUFFONE, GJ ;
SCHIMBOR, CM ;
MAY, RA .
JOURNAL OF INFECTIOUS DISEASES, 1988, 158 (06) :1177-1184
[8]   GENERATION OF SINGLE-STRANDED-DNA BY THE POLYMERASE CHAIN-REACTION AND ITS APPLICATION TO DIRECT SEQUENCING OF THE HLA-DQA LOCUS [J].
GYLLENSTEN, UB ;
ERLICH, HA .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1988, 85 (20) :7652-7656
[9]   DETECTION OF SERUM HEPATITIS-B VIRUS-DNA IN PATIENTS WITH CHRONIC HEPATITIS USING THE POLYMERASE CHAIN-REACTION ASSAY [J].
KANEKO, S ;
MILLER, RH ;
FEINSTONE, SM ;
UNOURA, M ;
KOBAYASHI, K ;
HATTORI, N ;
PURCELL, RH .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1989, 86 (01) :312-316
[10]  
LAWYER FC, 1989, J BIOL CHEM, V264, P6427