Cloning and development of synthetic internal amplification control for Bacillus anthracis real-time polymerase chain reaction assays

Youvraj Sohni; Sagarika Kanjilal; Vivek Kapur

doi:10.1016/j.diagmicrobio.2008.04.005

Cloning and development of synthetic internal amplification control for Bacillus anthracis real-time polymerase chain reaction assays

Youvraj Sohni, Sagarika Kanjilal, Vivek Kapur

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

9 Scopus citations

Abstract

An internal amplification control (IAC) was developed for Bacillus anthracis rpoB gene detection using TaqMan assay. Synthetic IAC oligonucleotides were subcloned using vector pDG1730 for ectopic integration into host Bacillus subtilis strain 1A772 genome. Differentially labeled target and IAC probes were used in real-time polymerase chain reaction (PCR) assays. There was no nonspecific cross-detection in single-well reactions. Limit of detection for both target and IAC DNA was 5 fg corresponding to a single gene copy. The IAC, in conjunction with target system, should decrease the rate of false-positive and false-negative results in real-time PCR assays.

Original language	English (US)
Pages (from-to)	471-475
Number of pages	5
Journal	Diagnostic Microbiology and Infectious Disease
Volume	61
Issue number	4
DOIs	https://doi.org/10.1016/j.diagmicrobio.2008.04.005
State	Published - Aug 2008

All Science Journal Classification (ASJC) codes

Microbiology (medical)
Infectious Diseases

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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10.1016/j.diagmicrobio.2008.04.005

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title = "Cloning and development of synthetic internal amplification control for Bacillus anthracis real-time polymerase chain reaction assays",

abstract = "An internal amplification control (IAC) was developed for Bacillus anthracis rpoB gene detection using TaqMan assay. Synthetic IAC oligonucleotides were subcloned using vector pDG1730 for ectopic integration into host Bacillus subtilis strain 1A772 genome. Differentially labeled target and IAC probes were used in real-time polymerase chain reaction (PCR) assays. There was no nonspecific cross-detection in single-well reactions. Limit of detection for both target and IAC DNA was 5 fg corresponding to a single gene copy. The IAC, in conjunction with target system, should decrease the rate of false-positive and false-negative results in real-time PCR assays.",

author = "Youvraj Sohni and Sagarika Kanjilal and Vivek Kapur",

note = "Funding Information: This research was supported in part by the US Department of Homeland Security (grant number N-00014-04-1-0659) through a grant awarded to the National Center for Food Protection and Defense at the University of Minnesota. Any opinions, findings, conclusions, or recommendations expressed in this publication are those of the author(s) and do not represent the policy or position of the Department of Homeland Security. Copyright: Copyright 2008 Elsevier B.V., All rights reserved.",

year = "2008",

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doi = "10.1016/j.diagmicrobio.2008.04.005",

language = "English (US)",

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journal = "Diagnostic Microbiology and Infectious Disease",

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AU - Kanjilal, Sagarika

AU - Kapur, Vivek

N1 - Funding Information: This research was supported in part by the US Department of Homeland Security (grant number N-00014-04-1-0659) through a grant awarded to the National Center for Food Protection and Defense at the University of Minnesota. Any opinions, findings, conclusions, or recommendations expressed in this publication are those of the author(s) and do not represent the policy or position of the Department of Homeland Security. Copyright: Copyright 2008 Elsevier B.V., All rights reserved.

PY - 2008/8

Y1 - 2008/8

N2 - An internal amplification control (IAC) was developed for Bacillus anthracis rpoB gene detection using TaqMan assay. Synthetic IAC oligonucleotides were subcloned using vector pDG1730 for ectopic integration into host Bacillus subtilis strain 1A772 genome. Differentially labeled target and IAC probes were used in real-time polymerase chain reaction (PCR) assays. There was no nonspecific cross-detection in single-well reactions. Limit of detection for both target and IAC DNA was 5 fg corresponding to a single gene copy. The IAC, in conjunction with target system, should decrease the rate of false-positive and false-negative results in real-time PCR assays.

AB - An internal amplification control (IAC) was developed for Bacillus anthracis rpoB gene detection using TaqMan assay. Synthetic IAC oligonucleotides were subcloned using vector pDG1730 for ectopic integration into host Bacillus subtilis strain 1A772 genome. Differentially labeled target and IAC probes were used in real-time polymerase chain reaction (PCR) assays. There was no nonspecific cross-detection in single-well reactions. Limit of detection for both target and IAC DNA was 5 fg corresponding to a single gene copy. The IAC, in conjunction with target system, should decrease the rate of false-positive and false-negative results in real-time PCR assays.

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Cloning and development of synthetic internal amplification control for Bacillus anthracis real-time polymerase chain reaction assays

Abstract

All Science Journal Classification (ASJC) codes

UN SDGs

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