Polymerase chain reaction detection of nonviable bacterial pathogens

K. L. Josephson, Charles P Gerba, Ian L Pepper

Research output: Contribution to journalArticle

318 Citations (Scopus)

Abstract

Polymerase chain reaction (PCR) methodologies for detection of pathogens in environmental samples are currently available. However, positive amplification products for any set of primers only signal that the appropriate target nucleic acid sequences were present in the sample. The presence of the amplification products does not imply that the target organisms were viable. Here we show that PCR will detect nonviable cells, as long as intact target nucleic acid sequences are available. In an environmental water sample, nucleic acids degraded quickly and were not detectable by PCR after 3 weeks even when stored at 4°C. However, these data show that there is a window of opportunity for PCR analyses to result in false positives with respect to viable cells. We further show that care must be taken in the way samples are stored for future PCR amplifications and that filter sterilization of media is not acceptable for long-term preservation of samples for PCR.

Original languageEnglish (US)
Pages (from-to)3513-3515
Number of pages3
JournalApplied and Environmental Microbiology
Volume59
Issue number10
StatePublished - 1993

Fingerprint

polymerase chain reaction
pathogen
Polymerase Chain Reaction
pathogens
nucleic acid
Nucleic Acids
nucleic acids
amplification
sampling
microbial detection
filters
detection
cells
filter
methodology
Water
organisms
water
product

ASJC Scopus subject areas

  • Biotechnology
  • Environmental Science(all)
  • Microbiology

Cite this

Polymerase chain reaction detection of nonviable bacterial pathogens. / Josephson, K. L.; Gerba, Charles P; Pepper, Ian L.

In: Applied and Environmental Microbiology, Vol. 59, No. 10, 1993, p. 3513-3515.

Research output: Contribution to journalArticle

@article{56e0bb39ece140788c67c6172ced1573,
title = "Polymerase chain reaction detection of nonviable bacterial pathogens",
abstract = "Polymerase chain reaction (PCR) methodologies for detection of pathogens in environmental samples are currently available. However, positive amplification products for any set of primers only signal that the appropriate target nucleic acid sequences were present in the sample. The presence of the amplification products does not imply that the target organisms were viable. Here we show that PCR will detect nonviable cells, as long as intact target nucleic acid sequences are available. In an environmental water sample, nucleic acids degraded quickly and were not detectable by PCR after 3 weeks even when stored at 4°C. However, these data show that there is a window of opportunity for PCR analyses to result in false positives with respect to viable cells. We further show that care must be taken in the way samples are stored for future PCR amplifications and that filter sterilization of media is not acceptable for long-term preservation of samples for PCR.",
author = "Josephson, {K. L.} and Gerba, {Charles P} and Pepper, {Ian L}",
year = "1993",
language = "English (US)",
volume = "59",
pages = "3513--3515",
journal = "Applied and Environmental Microbiology",
issn = "0099-2240",
publisher = "American Society for Microbiology",
number = "10",

}

TY - JOUR

T1 - Polymerase chain reaction detection of nonviable bacterial pathogens

AU - Josephson, K. L.

AU - Gerba, Charles P

AU - Pepper, Ian L

PY - 1993

Y1 - 1993

N2 - Polymerase chain reaction (PCR) methodologies for detection of pathogens in environmental samples are currently available. However, positive amplification products for any set of primers only signal that the appropriate target nucleic acid sequences were present in the sample. The presence of the amplification products does not imply that the target organisms were viable. Here we show that PCR will detect nonviable cells, as long as intact target nucleic acid sequences are available. In an environmental water sample, nucleic acids degraded quickly and were not detectable by PCR after 3 weeks even when stored at 4°C. However, these data show that there is a window of opportunity for PCR analyses to result in false positives with respect to viable cells. We further show that care must be taken in the way samples are stored for future PCR amplifications and that filter sterilization of media is not acceptable for long-term preservation of samples for PCR.

AB - Polymerase chain reaction (PCR) methodologies for detection of pathogens in environmental samples are currently available. However, positive amplification products for any set of primers only signal that the appropriate target nucleic acid sequences were present in the sample. The presence of the amplification products does not imply that the target organisms were viable. Here we show that PCR will detect nonviable cells, as long as intact target nucleic acid sequences are available. In an environmental water sample, nucleic acids degraded quickly and were not detectable by PCR after 3 weeks even when stored at 4°C. However, these data show that there is a window of opportunity for PCR analyses to result in false positives with respect to viable cells. We further show that care must be taken in the way samples are stored for future PCR amplifications and that filter sterilization of media is not acceptable for long-term preservation of samples for PCR.

UR - http://www.scopus.com/inward/record.url?scp=0027363920&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0027363920&partnerID=8YFLogxK

M3 - Article

C2 - 8250575

AN - SCOPUS:0027363920

VL - 59

SP - 3513

EP - 3515

JO - Applied and Environmental Microbiology

JF - Applied and Environmental Microbiology

SN - 0099-2240

IS - 10

ER -