Application of the method of multiplex polymerase chain reaction to identify the species composition of milk and dairy products

2021 ◽  
pp. 12-14
Author(s):  
Николай Анатольевич Жижин

Идентификация пищевого сырья, применяемого для производства продуктов питания, является важным аспектом безопасности здоровья человека. Развитие аллергических реакций, непереносимость отдельных компонентов пищи и т. д. являются поводом для вынесения информации о составе пищевой продукции на этикеточную надпись. Также к важным факторам оценки продуктов питания можно отнести и видовую «чистоту». Этот показатель необходимо учитывать не только в качестве выявления фальсификации продукции более дешевым сырьем, но и для безопасности конечного потребителя. Для идентификации видового состава молока и молочной продукции достаточно успешно используется метод полимеразной цепной реакции. Развитие этого метода привело к появлению различных подходов его использования. Одним из них является метод мультиплексной полимеразной реакции, который позволяет одновременно проводить процесс амплификации различных последовательностей ДНК. Что позволяет использовать данный метод для одновременного определения двух и более видов сельскохозяйственных животных в течение проведения одного ПЦР-анализа. В данной работе показаны возможности применения мультиплексной ПЦР для идентификации молока и молочной продукции. Показано, что в течение одного анализа определяются специфические праймеры, характерные для трех видов животных: коровы, овцы и козы. Применяемая методика также была использована на молочной продукции, прошедшей термическую обработку, в результате чего установлена возможность использования мультиплексной ПЦР для анализа такой продукции. Предел обнаружения при проведении ПЦР-анализа составил 0,1 %. The identification of food raw materials used for food production is an important aspect of human health safety. Development of allergic reactions, intolerance to certain food components, etc. are the reason for placing information on the composition of food products on the label inscription. Species «purity» can also be attributed to the important factors in assessing food products. This indicator must be taken into account not only as a detection of product counterfeiting with cheaper raw materials, but also for the safety of the end consumer. To identify the species composition of milk and dairy products, the method of polymerase chain reaction is quite successfully used. The development of this method has led to the emergence of various approaches to its use. One of which is the multiplex polymerase reaction method, which allows simultaneous amplification of various DNA sequences. That allows you to use this method for the simultaneous determination of two or more species of farm animals during one PCR analysis. In this work, the possibilities of using multiplex PCR for the identification of milk and dairy products were shown. It was shown that during one analysis, specific primers characteristic of three species of animals: cow, sheep and goat are determined. The applied technique was also used on heat-treated dairy products, as a result of which the possibility of using multiplex PCR for the analysis of such products was established. The detection limit for PCR analysis was 0.1 %.

2001 ◽  
Vol 64 (5) ◽  
pp. 664-668 ◽  
Author(s):  
SUDHIR TAMARAPU ◽  
JOHN L. McKILLIP ◽  
MARYANNE DRAKE

A multiplex polymerase chain reaction (PCR) assay was developed for the detection and differentiation of enterotoxigenic Staphylococcus aureus in dairy products. A solvent extraction procedure was successfully modified for extraction of S. aureus DNA from 10 ml of artificially contaminated skim milk or 20 g cheddar cheese. Primers targeting the enterotoxin C gene (entC) and thermostable nuclease gene (nuc) were used in the multiplex PCR. PCR products were confirmed using restriction fragment length polymorphism analysis. DNA was consistently quantified and amplified by uniplex PCR from 10 CFU/ml of S. aureus in skim milk or 10 CFU/20 g cheddar cheese. The sensitivity of the multiplex PCR was 100 CFU/ml of skim milk or 100 CFU/20 g cheddar cheese. The developed methodology allows presumptive identification and differentiation of enterotoxigenic S. aureus in less than 6 h.


LWT ◽  
2015 ◽  
Vol 63 (1) ◽  
pp. 714-719 ◽  
Author(s):  
Sahilah Abd Mutalib ◽  
Nursheila Mustafa Muin ◽  
Aminah Abdullah ◽  
Osman Hassan ◽  
Wan Aida Wan Mustapha ◽  
...  

Author(s):  
Jun-Hyung Lim ◽  
Sang Hwan Nam ◽  
Jongwoo Kim ◽  
Nam Hoon Kim ◽  
Gun-Soo Park ◽  
...  

Abstract In this study, a three-stage bioaerosol sampler with a sampling flow rate of 170 L/min was designed and fabricated for sampling the bioaerosols released during human breathing and coughing, and its performance was evaluated. The sampler was constructed using a cyclone separator with a cutoff size of 2.5 µm as a preseparator, a multi-nozzle virtual impactor with a cutoff size of 0.34 µm as an aerosol concentrator, and a BioSampler, which is a commercial product, for collecting bioaerosols in a collection fluid. The collection efficiency of the sampler was evaluated through simulations and experiments. Only particles with sizes of 0.1-4 µm were selectively collected in the collection fluid. Bacteriophage bioaerosols were sampled using the developed sampler and ACD-200 Bobcat sampler, which is a commercial product. The amounts of collected bacteriophages were compared using the polymerase chain reaction (PCR) technique. The sampling performance of the developed sampler was similar to that of the ACD-200 Bobcat sampler. Moreover, the developed sampler showed its ability to sample bioaerosols of a specific size-range and collect them directly in a collection fluid for the PCR analysis. Therefore, the developed sampler is expected to be useful for indoor environmental monitoring by effectively sampling the bioaerosols released indoors during human breathing and coughing.


2021 ◽  
Vol 15 ◽  
Author(s):  
Sara Galeb ◽  
Maysaa El Sayed Zaki ◽  
Raghdaa Shrief ◽  
Rasha Hassan ◽  
Mohamed Anies

Background: Proper identification of the causative organism in pediatric sepsis is crucial for early diagnosis and prevention of septic shock and organ failure. Objectives: The present study aimed to evaluate the multiplex Polymerase Chain Reaction (PCR) to detect Acinetobacter baumannii, Pseudomonas aeruginosa, and Stenotrophomonas maltophilia from positive blood cultures for these pathogens isolated from children, with hospital-acquired sepsis compared to the conventional biochemical reactions for identification of these organisms. Methods: This study was a cross-sectional study performed on 100 isolates from pediatric blood cultures, including Acinetobacter baumannii, Pseudomonas aeruginosa, and Stenotrophomonas maltophilia. The study also included 100 isolates of Escherichia coli as a negative control. All isolates were identified by API 20NE and the multiplex PCR, with primers specific to the 3 tested bacteria. Results: Multiplex PCR was positive in 96% of isolates, and 4 isolates had negative results. False positive results were reported with three E. coli strains. Multiplex PCR identified all the isolates of Acinetobacter baumannii, 29 isolates of Pseudomonas aeruginosa, and 27 isolates of Stenotrophomonas maltophilia. Compared to the biochemical identification, the diagnostic value of the multiplex PCR revealed 96.04% sensitivity, 96.9% specificity, 97.00%, positive predictive value, 96.00% negative predictive value, and 96.50% accuracy. Conclusion: The present study highlights the diagnostic value of multiplex PCR to identify Acinetobacter baumannii, Pseudomonas aeruginosa, and Stenotrophomonas maltophilia from positive blood cultures. Multiplex PCR was sensitive, specific, and accurate. The accuracy differs according to the organisms, with 100% accuracy for Acinetobacter baumannii.


2009 ◽  
Vol 27 (36) ◽  
pp. 6094-6100 ◽  
Author(s):  
Lindsey Goff ◽  
Karin Summers ◽  
Sameena Iqbal ◽  
Jens Kuhlmann ◽  
Michael Kunz ◽  
...  

Purpose The randomized First-Line Indolent Trial (FIT) was conducted in patients with advanced follicular lymphoma (FL), to evaluate the safety and efficacy of yttrium-90 (90Y) ibritumomab tiuxetan given as consolidation of complete or partial remission. This study of minimal residual disease was undertaken in parallel, to determine the rate of conversion from bcl-2 polymerase chain reaction (PCR) –detectable to –undetectable status and the corresponding effect on progression-free survival (PFS). Patients and Methods Blood samples from 414 patients (90Y-ibritumomab, n = 208; control, n = 206) were evaluated using real-time quantitative polymerase chain reaction (RQ-PCR); 186 were found to have the bcl-2 rearrangement and were thus eligible for inclusion in the RQ-PCR analysis. Results Overall, 90% of treated patients converted from bcl-2 PCR–detectable to –undetectable disease status, compared with 36% in the control group. Treatment significantly prolonged median PFS in patients converting to bcl-2 PCR-undetectable status (40.8 v 24.0 months in the control group; P < .01, hazard ratio [HR], 0.399). In patients who had bcl-2 PCR-detectable disease at random assignment, treatment significantly prolonged median PFS (38.4 v 8.2 months in the control group; P < .01, HR, 0.293). Conclusion Eradication of PCR-detectable disease occurred more frequently after treatment with 90Y-ibritumomab tiuxetan and was associated with prolongation of PFS.


2016 ◽  
Vol 7 (1) ◽  
Author(s):  
Celenk Molva ◽  
Halil Ibrahim Atabay

Arcobacters are food and waterborne pathogens associated with human and animal infections. The objective of the present study was to investigate the prevalence and diversity of <em>Arcobacter</em> spp. in commercially sold chicken meat in İzmir region of Turkey. For this purpose, 100 samples including legs (n=40), 17 chicken quarters (n=17), drumstickers (n=16), breasts (n=11), wings (n=10), and carcasses (n=6) were collected from different retail markets. A total of 65 isolates were confirmed as <em>Arcobacter</em> spp. from 55 samples by genus-specific polymerase chain reaction (PCR). The prevalence of <em>Arcobacter</em> spp. was 32.5, 81.3, 64.7, 72.7, 83.3, and 50% for legs, drumstickers, chicken quarters, breasts, carcasses and wings, respectively. Based on the multiplex-PCR, most of the isolates were identified as <em>A. butzleri</em> (n=45, 80%), followed by <em>A. cryaerophilus</em> (n=2, 3.6%), <em>A. skirrowii</em> (n=1, 1.8%) and 17 isolates (30.9%) could not be identified at the species level.


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