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Establishment of a Fluorescence-Based Quantitative Real-Time PCR Method for Detection for Caprine-Derived Ingredients in Milk Powder
CHEN Chen, ZHANG Rui, WANG Yuxin, ZHANG Tonghe, ZHANG Zilun, JIA Wenshen, ZHOU Wei
Journal of Dairy Science and Technology    2023, 46 (6): 20-24.   DOI: 10.7506/rykxyjs1671-5187-20231224-062
Abstract112)   HTML3)    PDF (1951KB)(61)       Save
This study established a fluorescence-based quantitative real-time polymerase chain reaction method for the detection of caprine-derived ingredients in milk powder. The specificity, sensitivity, and stability of the caprine-specific primers were evaluated. By linear fitting of the difference in cycle threshold (ΔCt) as a function of the mixing proportion between goat and horse milk powder, a calibration curve for the relative quantitation of caprine-derived ingredients in milk powder was established as follows: y = 0.720 9x + 5.651 9 (R2 = 0.987 5), and the minimum detection limit of this method was 0.000 1 ng/μL. The recoveries of the proposed method were 96.22%-112.00%, and the inter- and intra-group coefficient of variation was ≤ 0.79% and ≤ 1.77%, respectively.
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A Review of Technologies for Aflatoxin Detection in Milk and Dairy Products
YANG Fan, HUANG Jianhui, CHEN Chen, GUAN Shuhui, LIU Guangpeng, ZHANG Yan
Journal of Dairy Science and Technology    2023, 46 (3): 46-51.   DOI: 10.7506/rykxyjs1671-5187-20230331-015
Abstract127)   HTML13)    PDF (1813KB)(350)       Save
Aflatoxins (AFTs) is a group of secondary metabolites produced by Aspergillus flavus and Aspergillus parasiticus, which has stable physicochemical properties. Aflatoxin B1 (AFB1), aflatoxin B2 (AFB2), aflatoxin G1 (AFG1), aflatoxin G2 (AFG2), aflatoxin M1 (AFM1), and aflatoxin M2 (AFM2) are the common analogues of AFT, with AFB1 being the most toxic and widespread one. The International Agency for Research on Cancer (IARC) under the World Health Organization has classified AFB1 as a Group 1 carcinogen. When cows eat contaminated feedstuffs such as peanut, corn, rice, soybean and wheat, some of the AFTs are converted in the body into AFM1, and AFM2, which can exist in milk and dairy products. This paper compares the difference between Chinese and international limits for AFTs in foods, and summarizes the methods for detecting aflatoxin in milk and dairy products such as thin-layer chromatography (TLC), mass spectrometry (MS), spectroscopy, electrochemistry, and rapid test strips. This paper analyzes the advantages and limitations of these methods and predicts future directions in the development of aflatoxin detection technology in milk and dairy products. It is hoped that this review will help in the development of a more convenient, specific, and sensitive detection method for aflatoxin in milk and dairy products.
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Establishment of a Helicase-Dependent Isothermal DNA Amplification Method for Rapid Detection of Lactobacillus plantarum in Fermented Milk
ZHANG Yalun, LI Yongbo, ZHANG Tao, CHEN Chen, ZHANG Jie, ZHOU Wei, ZHANG Yan
Journal of Dairy Science and Technology    2023, 46 (1): 30-34.   DOI: 10.7506/rykxyjs1671-5187-20220908-056
Abstract24)   HTML7)    PDF (2026KB)(20)       Save
A rapid detection method based on helicase-dependent isothermal DNA amplification (HDA) was established for Lactobacillus plantarum in fermented milk. Specific primers were designed according to the scrB gene sequence of Lactobacillus plantarum (Genbank Accession NO. AJ579541.1), and the optimal concentrations of UvrD helicase and T4 gp32 in the reaction system were determined through experiments. The limit of detection (LOD), specificity, consistency and stability of the proposed method were evaluated by use of L. plantarum-spiked samples, amplification of various strains, and electrophoresis of amplified products and sequence alignment analysis, respectively. The results showed that the optimized of UvrD helicase and T4 gp32 in the reaction system were found to be 0.15 and 5.0 μg, respectively. The HDA method had high specificity with no amplification of other strains tested. The detection limit was 2.8 × 101 CFU/g. The amplified product was consistent with the designed sequence length (273 bp) and the sequence homology was 100%. In conclusion: this method is rapid, simple, sensitive and suitable for the detection of Lactobacillus plantarum in fermented milk.
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Development of Ready-to-Use Quality Control Samples for the Detection of Salmonella in Formula Foods for Special Medical Purposes Containing Milk
ZHANG Tao, ZHANG Rui, ZHANG Yalun, XU Miaomiao, WANG Hui, CHEN Chen, ZHOU Wei, SHI Guohua
Journal of Dairy Science and Technology    2021, 44 (4): 29-33.   DOI: 10.15922/j.cnki.jdst.2021.04.007
Abstract156)   HTML0)    PDF (1990KB)(149)       Save
In order to evaluate the application value of ready-to-use Salmonella quality control samples in matrix assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) identification of Salmonella in milk-containing formula foods for special medical purposes, this experiment evaluated the uniformity and stability of the quality control samples. After being stored at 20 ℃ for 0, 14 and 28 days, these samples were identified by mass spectrometry and the VITEK automatic microbial analysis system. The results showed that the quality control samples had good stability and uniformity, and their mass spectra were similar to each other. The VITEK system confirmed their identity as Salmonella. In conclusion, the ready-to-use Salmonella quality control strains can be used for the detection of Salmonella in milk-containing formula foods for special medical purposes.
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