An RNA transcription-based amplification technique (NASBA) for the detection of viable Salmonella enterica

S. A. Simpkins; A. B. Chan; J. Hays; B. Pöpping; N. Cook

doi:10.1046/j.1472-765x.2000.00670.x

An RNA transcription-based amplification technique (NASBA) for the detection of viable Salmonella enterica

S. A. Simpkins, A. B. Chan, J. Hays, B. Pöpping, N. Cook^*

^*Corresponding author for this work

Research output: Contribution to journal › Article › Academic › peer-review

112 Citations (Scopus)

6 Downloads (Pure)

Abstract

Possession of mRNA is indicative of cell viability. RTPCR is not appropriate for mRNA detection as it cannot unambiguously detect mRNA in a DNA background. The alternative amplification technique, NASBA, avoids the disadvantages of RTPCR. We have devised a method for detection of viable Salmonella enterica. This involves NASBA amplification of mRNA transcribed from the dnaK gene. Amplification of mRNA extracted from viable and heat-killed cells from the same population produced consistent and highly significant (P > 0 01) differences between the respective signals. The signal obtained from viable cells was completely eradicated by RNase treatment, while PCR amplification of treated and untreated samples was unaffected, indicating that NASBA was unaffected by background DNA.

Original language	English
Pages (from-to)	75-79
Number of pages	5
Journal	Letters in Applied Microbiology
Volume	30
Issue number	1
DOIs	https://doi.org/10.1046/j.1472-765x.2000.00670.x
Publication status	Published - 2000
Externally published	Yes

Access to Document

10.1046/j.1472-765x.2000.00670.xLicence: Article 25fa Dutch Copyright Act

An RNA transcription-based amplification technique (NASBA) for the detection of viable Salmonella entericaFinal published version, 128 KBLicence: Article 25fa Dutch Copyright Act

Cite this

@article{adc61910b93a4f5ba67e9068db4683ab,

title = "An RNA transcription-based amplification technique (NASBA) for the detection of viable Salmonella enterica",

abstract = "Possession of mRNA is indicative of cell viability. RTPCR is not appropriate for mRNA detection as it cannot unambiguously detect mRNA in a DNA background. The alternative amplification technique, NASBA, avoids the disadvantages of RTPCR. We have devised a method for detection of viable Salmonella enterica. This involves NASBA amplification of mRNA transcribed from the dnaK gene. Amplification of mRNA extracted from viable and heat-killed cells from the same population produced consistent and highly significant (P > 0 01) differences between the respective signals. The signal obtained from viable cells was completely eradicated by RNase treatment, while PCR amplification of treated and untreated samples was unaffected, indicating that NASBA was unaffected by background DNA.",

author = "Simpkins, {S. A.} and Chan, {A. B.} and J. Hays and B. P{\"o}pping and N. Cook",

year = "2000",

doi = "10.1046/j.1472-765x.2000.00670.x",

language = "English",

volume = "30",

pages = "75--79",

journal = "Letters in Applied Microbiology",

issn = "0266-8254",

publisher = "Wiley-Blackwell Publishing Ltd",

number = "1",

}

TY - JOUR

T1 - An RNA transcription-based amplification technique (NASBA) for the detection of viable Salmonella enterica

AU - Simpkins, S. A.

AU - Chan, A. B.

AU - Hays, J.

AU - Pöpping, B.

AU - Cook, N.

PY - 2000

Y1 - 2000

N2 - Possession of mRNA is indicative of cell viability. RTPCR is not appropriate for mRNA detection as it cannot unambiguously detect mRNA in a DNA background. The alternative amplification technique, NASBA, avoids the disadvantages of RTPCR. We have devised a method for detection of viable Salmonella enterica. This involves NASBA amplification of mRNA transcribed from the dnaK gene. Amplification of mRNA extracted from viable and heat-killed cells from the same population produced consistent and highly significant (P > 0 01) differences between the respective signals. The signal obtained from viable cells was completely eradicated by RNase treatment, while PCR amplification of treated and untreated samples was unaffected, indicating that NASBA was unaffected by background DNA.

AB - Possession of mRNA is indicative of cell viability. RTPCR is not appropriate for mRNA detection as it cannot unambiguously detect mRNA in a DNA background. The alternative amplification technique, NASBA, avoids the disadvantages of RTPCR. We have devised a method for detection of viable Salmonella enterica. This involves NASBA amplification of mRNA transcribed from the dnaK gene. Amplification of mRNA extracted from viable and heat-killed cells from the same population produced consistent and highly significant (P > 0 01) differences between the respective signals. The signal obtained from viable cells was completely eradicated by RNase treatment, while PCR amplification of treated and untreated samples was unaffected, indicating that NASBA was unaffected by background DNA.

UR - http://www.scopus.com/inward/record.url?scp=0034010361&partnerID=8YFLogxK

U2 - 10.1046/j.1472-765x.2000.00670.x

DO - 10.1046/j.1472-765x.2000.00670.x

M3 - Article

C2 - 10728566

AN - SCOPUS:0034010361

SN - 0266-8254

VL - 30

SP - 75

EP - 79

JO - Letters in Applied Microbiology

JF - Letters in Applied Microbiology

IS - 1

ER -

An RNA transcription-based amplification technique (NASBA) for the detection of viable Salmonella enterica

Abstract

Access to Document

Other files and links

Fingerprint

Cite this