Detection of Cronobacter sakazakii in Infant Formula Powder by Helicase-Dependent Isothermal DNA Amplification Assay

doi:10.7506/spkx1002-6630-201404032

Abstract

Abstract:

Objective: The aim of this study was to establish a helicase-dependent isothermal DNA amplification
(HAD) method for rapid and accurate detection of Cronobacter sakazakii in infant formula powder. Methods: A pair of
oligonucleotide primers exclusively targeting the internal transcribed spacer (ITS) gene of Cronobacter sakazakii was
designed, and the final concentration of UvrD helicase and T4 gp32 were optimized. Cronobacter sakazakii in infant formula
powder was then directly detected by this method and amplification products were separated and detected by agarose gel
electrophoresis. Results: An amplicon of 100 bp in length was obtained, which was the same as the designed gene fragment
by HDA. The sensitivity of HDA was 101 CFU/g, the optimized concentration of UvrD helicase was 0.1 μg, and T4 gp32
was 5.0 μg. Conclusion: The helicase-dependent isothermal DNA amplification assay provides a new, specific, sensitive and
fast for the detection of Cronobacter sakazakii in infant formula powder.

Key words: helicase-dependent isothermal DNA amplification (HDA), infant formula powder, Cronobacter sakazakii, detection

CLC Number:

TS201.3

ZHOU Wei1,2, ZHANG Wei2, LIU Liang2, LIU Dong2, LI Yong-bo2, TIAN Hao2, ZHANG Yan2,*, ZHANG Zhi-sheng1,*. Detection of Cronobacter sakazakii in Infant Formula Powder by Helicase-Dependent Isothermal DNA Amplification Assay[J]. FOOD SCIENCE, doi: 10.7506/spkx1002-6630-201404032.

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Detection of Cronobacter sakazakii in Infant Formula Powder by Helicase-Dependent Isothermal DNA Amplification Assay

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