Development of gold nanoparticle-based visual assay for rapid detection of Escherichia coli specific DNA in milk of cows affected with mastitis

For early diagnosis of infections, a rapid, sensitive and precise method to identify microbial DNA coming from veterinary clinical samples, such as milk of cows affected with mastitis, is fascinating. We developed an amplification-free visual assay for rapid and sensitive detection of specific DNA f...

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Bibliographic Details
Published inFood science & technology Vol. 155; p. 112901
Main Authors Deb, Rajib, Pal, Prasanna, Chaudhary, Parul, Bhadsavle, Sanat, Behera, Manisha, Parmanand, Gautam, Devika, Roshan, Mayank, Vats, Ashutosh, Ludri, Ashutosh, Gupta, Vivek Kumar, De, Sachinandan
Format Journal Article
LanguageEnglish
Published Elsevier Ltd 01.02.2022
Subjects
Amplification-free
Escherichia coli
Gold nanoparticles
uid A
Visual assay
LOD
uid A
AC
SE
CFU
Visual assay
Gold nanoparticles
Escherichia coli
AuNPs
uidA
SP
Amplification-free
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Summary:For early diagnosis of infections, a rapid, sensitive and precise method to identify microbial DNA coming from veterinary clinical samples, such as milk of cows affected with mastitis, is fascinating. We developed an amplification-free visual assay for rapid and sensitive detection of specific DNA from Escherichia coli, particularly uid A gene encoding for beta-glucuronidase, isolated in milk of cows affected with mastitis, based on multiple gold nanoparticles (AuNPs) captured on a magnetic microbead surface, leading to plasmonic signal enhancement, thereby improving overall sensitivity of the assay. This test can be performed in 1–1.5 h after post template DNA preparation and can be visualized with naked eyes without the use of any expensive equipment. The visual assay detects a minimum microbial load of 102 CFU/μl and can identify bacterial DNA as low as 1 pg. This test provides a precise detection tool for E coli in clinical samples, such as milk of cows affected with mastitis, as a quick and user-friendly molecular detection approach. •Developed visual assay can specifically detect Escherichia coli DNA in clinical mastitis milk samples.•The assay can detect bacterial DNA as low as 102 CFU/μl.•The assay can be visualized in naked eyes and can be performed without any costly equipment.
ISSN:0023-6438
1096-1127
DOI:10.1016/j.lwt.2021.112901