A rapid and efficient DNA isolation method for qPCR-based detection of pathogenic and spoilage bacteria in milk

被引：10

作者：

Pancza, Bettina ^{[1
]}

Szathmary, Miklos ^{[1
]}

Gyurjan, Istvan ^{[2
]}

Bankuti, Barbara ^{[3
]}

Tudos, Zoltan ^{[2
]}

Szathmary, Susan

Stipkovits, Laszlo ^{[1
]}

Sipos-Kozma, Zsofia ^{[2
]}

Asvanyi, Balazs ^{[2
]}

Varga, Laszlo ^{[4
]}

Szenthe, Kalman ^{[1
]}

Banati, Ferenc ^{[3
]}

机构：

[1] Carlsbad Res Org, 2 Szabadsag St, H-9244 Ujronafo, Hungary

[2] Galen Bio Ltd, 15-17 Lucsony St, H-9200 Mosonmagyarovar, Hungary

[3] RT Europe Nonprofit Res Ctr, 2 Var Sq,Bldg E, H-9200 Mosonmagyarovar, Hungary

[4] Szechenyi Istvan Univ, Fac Agr & Food Sci, Dept Food Sci, 15-17 Lucsony St, H-9200 Mosonmagyarovar, Hungary

来源：

FOOD CONTROL | 2021年 / 130卷

关键词：

Raw milk; Pathogenic bacteria; DNA purification; Pretreatment; qPCR; CHAIN-REACTION PCR; REAL-TIME PCR; BOVINE-MILK; EXTRACTION;

D O I：

10.1016/j.foodcont.2021.108236

中图分类号：

TS2 [食品工业];

学科分类号：

0832 ;

摘要：

The objective of this study was to find an efficient, rapid, simple, and cost-effective method of pretreating raw milk samples to produce PCR-ready DNA for subsequent microbial detection using the strains of eight bacterial species. A total of 17 in-house protocols and three commercial kits were evaluated in three steps from scientific, practical, and economic perspectives. The results showed that an in-house procedure involving Triton X-100 based pretreatment and an inhibitor removal resin was superior to all other methods tested in terms of DNA yield, sensitivity, ease of sample handling, time efficiency, and cost per sample. Overall, this simplified pre analytical protocol was shown to have a great potential for use in rapid detection of dairy-related bacterial species, thereby enabling early intervention in the food chain and thus reducing the risk of negative economic and health outcomes.

引用

页数：9

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