Biotechnical use of polymerase chain reaction for microbiological analysis of biological samples

Biotechnol Annu Rev. 2000;5:87-130. doi: 10.1016/s1387-2656(00)05033-x.

Abstract

Since its introduction in the mid-80s, polymerase chain reaction (PCR) technology has been recognised as a rapid, sensitive and specific molecular diagnostic tool for the analysis of micro-organisms in clinical, environmental and food samples. Although this technique can be extremely effective with pure solutions of nucleic acids, it's sensitivity may be reduced dramatically when applied directly to biological samples. This review describes PCR technology as a microbial detection method, PCR inhibitors in biological samples and various sample preparation techniques that can be used to facilitate PCR detection, by either separating the micro-organisms from PCR inhibitors and/or by concentrating the micro-organisms to detectable concentrations. Parts of this review are updated and based on a doctoral thesis by Lantz [1] and on a review discussing methods to overcome PCR inhibition in foods [2].

Publication types

  • Research Support, Non-U.S. Gov't
  • Review

MeSH terms

  • Animals
  • DNA / isolation & purification*
  • DNA Primers
  • DNA-Directed DNA Polymerase / chemistry
  • DNA-Directed DNA Polymerase / physiology
  • Enzyme Inhibitors / metabolism
  • Enzyme Inhibitors / pharmacology
  • Humans
  • Microbiological Techniques*
  • Nucleic Acid Synthesis Inhibitors
  • Polymerase Chain Reaction / methods*
  • Reverse Transcriptase Polymerase Chain Reaction / methods
  • Specimen Handling / methods*

Substances

  • DNA Primers
  • Enzyme Inhibitors
  • Nucleic Acid Synthesis Inhibitors
  • DNA
  • DNA-Directed DNA Polymerase