Development and validation of fast Real-Time PCR assays for species identification in raw and cooked meat mixtures

Cammà, Cesare; Domenico, Marco Di; Monaco, Federica

doi:10.1016/j.foodcont.2011.08.007

Cited by 96 publications

(58 citation statements)

References 27 publications

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“…The concentration of genomic DNA was measured with a Nanodrop spectrophotometer (Thermo Fisher Scientific Inc., Wilmington, DE, USA) and Q-PCR was performed in triplicate in order to create the standard curve. The amplification efficiency (AE) of Q-PCR was calculated following the formula E = (10 -1/slope -1) × 100 [5]. Lower AE means that the quantity of PCR products generated at each cycle decreases so that the amplification will be take longer.…”

Section: Limit Of Detection (Lod) Of Q-pcrmentioning

confidence: 99%

“…Although the LOD of the Q-PCR was one order lower than that of the ME biosensor, the LOD is a possible limiting factor for either method, especially when Salmonella contamination may occur with 1000 cells or less. Nonetheless, the fact that the LOD of the ME biosensor was only one order higher than that of the Q-PCR showed that the ME biosensor method is competitive to Q-PCR, considered to be one of the most sensitive detection methods known to date [5,11,16].…”

Section: Limit Of Detection (Lod) Of Me Biosensormentioning

confidence: 99%

“…Considerable effort has been directed towards the development of rapid and simple detection methods such as polymerase chain reaction (PCR), enzyme-linked immunosorbent assay (ELISA), and several types of biosensor methods in order to substitute for the laborious and time-consuming conventional detection methods [5,29,30]. PCR is one of most promising alternative methods of detecting Salmonella in foods since many studies have shown that PCR is well suited for rapid, sensitive, specific and accurate bimolecular detection [5,11,16].…”

Section: Introductionmentioning

confidence: 99%

“…PCR is one of most promising alternative methods of detecting Salmonella in foods since many studies have shown that PCR is well suited for rapid, sensitive, specific and accurate bimolecular detection [5,11,16]. In addition, recently emerging PCR assays with fluorogenic probes (dye SYBR green, TaqMan, molecular beacons, and scorpion) have facilitated the development of real-time PCR assays more accurate and sensitive towards the identification and quantification for Salmonella [5,8,20]. In a preliminary study, a TaqMan-based quantitative real-time PCR method (Q-PCR) for the detection of S. typhimurium on tomato surfaces was demonstrated.…”

Section: Introductionmentioning

confidence: 99%

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Comparison of Phage-Based Magnetoelastic Biosensors with Taqman- Based Quantitative Real-Time PCR for the Detection of Salmonella typhimurium Directly Grown on Tomato Surfaces

Park

Park²,

Wikle³

et al. 2012

J Biosens Bioelectron

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A phage-based magnetoelastic (ME) biosensor method was compared with a TaqMan-based quantitative realtime PCR (Q-PCR) method for the detection of Salmonella typhimurium on tomato surfaces. This ME biosensor method utilizes magnetoelastic resonators coated with E2 filamentous phage to bind with and measure the concentration of S. typhimurium. In this study, standard curves, correlations, and limits of detection (LOD) for the ME biosensor and Q-PCR methods were determined by inoculating tomato surfaces with S. typhimurium suspensions in concentrations ranging from 1 to 8 log CFU/tomato. The LOD for the ME biosensor method and Q-PCR were 3 and 2 log CFU/tomato, respectively. In a direct comparison of the detection methods, S. typhimurium suspensions (3 log CFU/tomato) were inoculated on 65 tomato surfaces, then incubated at 37°C and 100% RH for 24 h. After 24 h, S. typhimurium was positively detected by both methods and the quantified concentrations were nearly the same, (6.35 ± 2.03) and (6.34 ± 0.17) log CFU/tomato respectively for the ME biosensor method and the Q-PCR method, which were significantly greater than the concentration determined by the BGS-plate count method (5.33 ± 0.21). Scanning electron microscopy (SEM) was used to confirm the growth of S. typhimurium on the tomato surfaces and the binding of S. typhimurium on the measurement sensors. This study demonstrated that the ME biosensor method was robust and competitive with Q-PCR for S. typhimurium detection on fresh produce.

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Section: Limit Of Detection (Lod) Of Q-pcrmentioning

confidence: 99%

Section: Limit Of Detection (Lod) Of Me Biosensormentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Comparison of Phage-Based Magnetoelastic Biosensors with Taqman- Based Quantitative Real-Time PCR for the Detection of Salmonella typhimurium Directly Grown on Tomato Surfaces

Park

Park²,

Wikle³

et al. 2012

J Biosens Bioelectron

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“…Determination of fraud in meat production is not only important for economic, health and ethical reasons but also to ensure fair trade and compliance with legislation (17,23). Due to several reasons including food scandals and socio-economic changes, consumers are demanding increase in the detection of meat species and fraudulent labeling in different foods (5). Also, authentication has become more important for Muslim countries in recent years, because of halal food status (21).…”

Section: Introductionmentioning

confidence: 99%

Identification of meat species in different types of meat products by PCR

Keyvan¹,

Çil²,

Kul³

et al. 2017

Ankara Üniversitesi Veteriner Fakültesi Dergisi

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Summary:Species identification in food of animal origin is an important subject for food control. Adulteration in foods is relevant for economical, religous, legislation or public health concerning reasons. The aim of this study was to determine the adulteration in different meat products such as sucuk (n=37), sausage (n=33) and salami (n=32). A total of 102 different meat products obtained from various markets in Ankara were analyzed for species identification by PCR. According to the analyzes, five (13.5%) poultry, and one (2.7%) poultry and equine meats together were found in the sucuk samples that are not declared in their labels. Also, seven (21.8%) and two (6.1%) poultry meat were detected in 32 salami and 33 sausage samples, respectively. These results indicate that 15 (14.7%) of the total samples were found to contain undeclared species. As a result, there were meat products which were not in compliance with their labels in various markets, presenting a potential public health risk and economical losses of consumers.Keywords: Adulteration, meat species identification, mislabeling, PCR. Farklı tip et ürünlerinde PZR ile tür tayiniÖzet: Et ürünlerinde tür tayini gıda güvenliği açısından önemli bir konudur. Etikette bildirilmeyen çeşitli hayvan türlerine ait etlerin kullanımı, tüketici sağlığı, ülke ekonomisi, dini ve yasal düzenlemeler ile ilişkilendirilmektedir. Bu çalışmada, sucuk, salam, sosis gibi ürünlerde farklı türlere ait etlerin varlığının tespiti amaçlanmıştır. Araştırmada Ankara'da çeşitli marketlerde satışa sunulan toplam 102 adet işlenmiş et ürünü tür tayini amacıyla PZR yöntemi kullanılarak incelenmiştir. Yapılan analizler sonucunda, incelenen 37 adet sucuk örneğinden etikette belirtilmeyen 5 (%13.5) kanatlı, 1 (%2.7) kanatlı ve tek tırnaklı eti tespit edilmiştir. Benzer şeklide, 32 salam örneğinin yedisinde (%21.8), 33 adet sosis örneğinin de ikisinde (%6.1) kanatlı eti bulunmuştur. Ürünlerin etiket bilgileri karşılaştırıldığında 15 adet (% 14.7) örneğin etiketinde belirtilmeyen farklı hayvan türlerine ait etlerden üretildiği tespit edilmiştir. Sonuç olarak, Ankara'da farklı marketlerden alınan işlenmiş et ürünleri örneklerinde, etiketlerinde belirtilmeyen hayvan türlerine ait etlerin kullanıldığı ve bu durumun potansiyel halk sağlığı sorunları yanında tüketicinin ekonomik olarak da kayba uğramasına sebep olabileceği ortaya konmuştur. Anahtar sözcükler: Hile, et tür tayini, yanlış etiketleme, PZR.

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