The development of a hexaplex-conventional PCR for identification of six animal and plant species in foodstuffs

Safdar, Muhammad; Junejo, Yasmeen

doi:10.1016/j.foodchem.2015.07.082

Cited by 31 publications

(16 citation statements)

References 29 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…However, meat adulteration affects food safety, quality, and many other respects, becoming a public focus recently. Since the 1980s, many immunological and molecular methods for species identification in food products have been developed [ 1 – 8 ]. Concomitant with advances in large-scale integrated molecular technology, DNA analysis methods have been in rapid development.…”

Section: Introductionmentioning

confidence: 99%

A PCR Method That Can Be Further Developed into PCR-RFLP Assay for Eight Animal Species Identification

Gao

Jin

Zhao

et al. 2018

Journal of Analytical Methods in Chemistry

View full text Add to dashboard Cite

There are many PCR-based methods for animal species identification; however, their detection numbers are limited or could not identify unknown species. We set out to solve this problem by developing a universal primer PCR assay for simultaneous identification of eight animal species, including goat, sheep, deer, buffalo, cattle, yak, pig, and camel. In this assay, the variable lengths of mitochondrial DNA were amplified using a pair of universal primers. PCR amplifications yielded 760 bp, 737 bp, 537 bp, 486 bp, 481 bp, 464 bp, 429 bp, and 359 bp length fragments for goat, sheep, deer, buffalo, cattle, yak, pig, and camel, respectively. This primer pair had no cross-reaction with other common domestic animals and fish. The limit of detection varied from 0.01 to 0.05 ng of genomic DNA for eight animal species in a 20 µl PCR mixture. Each PCR product could be further digested into fragments with variable sizes and qualitative analysis by SspI restriction enzyme. This developed PCR-RFLP assay was sufficient to distinguish all targeted species. Compared with the previous published related methods, this approach is simple, with high throughput, fast processing rates, and more cost-effective for routine identification of meat in foodstuffs.

show abstract

Section: Introductionmentioning

confidence: 99%

A PCR Method That Can Be Further Developed into PCR-RFLP Assay for Eight Animal Species Identification

Gao

Jin

Zhao

et al. 2018

Journal of Analytical Methods in Chemistry

View full text Add to dashboard Cite

show abstract

“…While previous studies reported an absolute LOD of 0.1 ng using multiplex PCR for horse, dog and pork 31 and 0.01 ng using real time PCR for horse 8,32 . (Safdar and Junejo 2016) 14 reported similar analytical sensitivities, namely 0.01% sensitivity for horse, soybean, sheep, poultry, pork, and cow, respectively. Therefore, the detection limit of the real-time PCR assay established in this study was equal or lower to previously reported values.…”

Section: Sensitivity Of the Method Considering The European Network mentioning

confidence: 85%

“…The most widely used identification method is based on DNA and a variety of techniques have been developed, including species-specific PCR (Polymerase Chain Reaction) [14][15][16] , PCR-RFLP (Polymerase Chain Reaction-Restriction Fragment Length Polymorphism) 9 , real-time PCR 17 , DNA barcoding 18 . Among them, a probe-based TaqMan ® real-time PCR is a robust assay with great rapidity and sensitivity.…”

Section: Development Of a Real-time Pcr Assay For The Identification mentioning

confidence: 99%

Development of a real-time PCR assay for the identification and quantification of bovine ingredient in processed meat products

Chen

Lü

Xiong

et al. 2020

Sci Rep

View full text Add to dashboard Cite

In order to find fraudulent species substitution in meat products, a highly sensitive and rapid assay for meat species identification and quantification is urgently needed. In this study, species-specific primers and probes were designed from the mitochondrial cytb (cytochrome b) fragment for identification and quantification of bovine ingredient in commercial meat products. Bovine samples and non-bovine ones were used to identify the specificity, sensitivity, and applicability of established assay. Results showed that the primers and probes were highly specific for bovine ingredient in meat products. The absolute detection limit of the real-time PCR method was 0.025 ng DNA, and the relative detection limit was 0.002% (w/w) of positive samples. The quantitative real-time PCR assay was validated on simulated meat samples and high in the precision and accuracy. In order to demonstrate the applicability and reliability of the proposed assay in practical products, the 22 commercial meat products including salted, jerkies, and meatball were used. The results indicated the established method has a good stability in detection of bovine ingredient in real food. The established method in this study showed specificity and sensitivity in identification and quantification of beef meat in processed meat products.

show abstract

“…Accurate labelling of food ingredients in meat products is a global issue because some individuals may have allergies, be lactose intolerance or have religious beliefs that require the avoidance of certain meats; for example, followers of Hinduism do not eat beef, and pork and horse are avoided by Muslims (Bottero & Dalmasso, ; Safdar & Junejo, ). In recognition of these concerns, the United States Food and Drug Administration (FDA) defined Economically Motivated Adulteration (EMA) of foods as the intentional adulteration of food for financial advantage, and the importance of food control and regulation to protect consumers from incorrect information about products’ components has been emphasised by several researchers (Everstine et al ., ; Strayer et al ., ).…”

Section: Introductionmentioning

confidence: 99%

“…Real‐time PCR method not only gives more simplicity and accuracy than conventional PCR but also has been used for quantitative detection. However, it is difficult to apply this method in most laboratories because the equipment and reagents are expensive and high technical skill is required (Safdar & Junejo, ). Recently, isothermal amplification method such as loop‐mediated isothermal amplification (LAMP) has been applied to identify meat species in processed meat products (Cho et al ., ; Kim & Kim, ).…”

Section: Introductionmentioning

confidence: 99%

Development and validation of a multiplex PCR assay for simultaneous detection of chicken, turkey and duck in processed meat products

Kim

Yoo

Yang

et al. 2018

Int J of Food Sci Tech

View full text Add to dashboard Cite

Summary We developed a multiplex PCR assay for the efficient detection of chicken, turkey and duck meat. Three species‐specific primer sets targeting 12S rRNA gene of turkey and mitochondrial cytochrome b (CYTB) gene of chicken and duck were selected. A universal primer pair targeting 18S rRNA gene was additionally used as a positive control. PCR product sizes for endogenous control, chicken, turkey and duck were confirmed to be 99 bp, 133 bp, 163 bp and 204 bp respectively. The specificity of each primer pairs was evaluated in 20 animal species. The limit of detection of this multiplex PCR assay was 1 pg for all species. Furthermore, this method was successfully applied to 31 commercial meat samples and validated at intra‐laboratory. This multiplex PCR assay for the simultaneous identification of chicken, turkey and duck meat in processed products can be utilised to monitor various processed food products for food control and regulation.

show abstract

The development of a hexaplex-conventional PCR for identification of six animal and plant species in foodstuffs

Cited by 31 publications

References 29 publications

A PCR Method That Can Be Further Developed into PCR-RFLP Assay for Eight Animal Species Identification

A PCR Method That Can Be Further Developed into PCR-RFLP Assay for Eight Animal Species Identification

Development of a real-time PCR assay for the identification and quantification of bovine ingredient in processed meat products

Development and validation of a multiplex PCR assay for simultaneous detection of chicken, turkey and duck in processed meat products

Contact Info

Product

Resources

About