DNA-based detection of pork content in food

Muflihah,; Hardianto, Ari; Kusumaningtyas, Pintaka; Prabowo, Sulistyo; Hartati, Yeni Wahyuni

doi:10.1016/j.heliyon.2023.e14418

Cited by 10 publications

(3 citation statements)

References 96 publications

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“…This initial cut is referred to as the "cis-activity" of Cas12a. In the second step, Cas12a indiscriminately degrades the ssDNA in a process known as "trans-activity" This trans-activity allows Cas12a to degrade not only the target DNA but also any other single-stranded DNA present in the system [26].…”

Section: Homology Difference Analysis Of the Selected Grnamentioning

confidence: 99%

Identification of CRISPR/Cas12a (Cpf1) guideRNA Sequence Targeting the Mitochondrial DNA D-loop Region in Wild Pig (Sus scrofa) Through Homology Difference and Mismatch Analysis

Zein,

Hardianto,

Irkham

et al. 2024

Trends Sci

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CRISPR (Clustered Regularly Interspaced Short Palindromic Repeat) or CRISPR-associated (Cas) System has become a major gene editing tool. Gene editing with CRISPR requires the Cas protein and the corresponding guide RNA (gRNA). However, low cleavage efficiency and off-target effects can impede the application of the CRISPR/Cas system. Therefore, the determination of specific gRNAs is essential. In biosensor applications, CRISPR/Cas12a can enhance specificity and sensitivity in identifying target genes due to the trans-cleavage activity of Cas12a (Cpf1). The mtDNA D-loop sequence is the most variable part of mtDNA, making it suitable for distinguishing between species. Consequently, the objective of this study was to determine the gRNA sequence of the D-loop of wild pig mtDNA in silico. Candidate gRNAs were predicted using the Benchling application with the assistance of the GenBank database. The gRNA candidates were subsequently subjected to a homology difference analysis using BLAST nucleotide and a mismatch test using Jalview. Among several candidates, candidate 1 was selected as the best option, with an Off-target value of 99.8. The homology difference analysis against competitors and the mismatch test against the Sus genus resulted in high E-values and high percentage values, respectively. This suggests that the candidate will not recognize other species but can detect members of the Sus scrofa species. These gRNA candidates can be selectively and sensitively applied to biosensors for the detection of meat adulteration. HIGHLIGHTS The D-loop exhibits high variability, make it useful for distinguishing between species gRNA as an enabler of CRISPR/Cas12a to identify the target and initiate cleavage activity The candidate 1 was selected with the sequence 5'-GAT TGT CGT GCC GGA TCA TGA GTT-3' The application of this strategy is aimed at determining the halal status of food products The application will encompass both quantitative and qualitative aspects GRAPHICAL ABSTRACT

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Section: Homology Difference Analysis Of the Selected Grnamentioning

confidence: 99%

Identification of CRISPR/Cas12a (Cpf1) guideRNA Sequence Targeting the Mitochondrial DNA D-loop Region in Wild Pig (Sus scrofa) Through Homology Difference and Mismatch Analysis

Zein,

Hardianto,

Irkham

et al. 2024

Trends Sci

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“…Homepage: https://journal.uii.ac.id/IJCA flexible use, fast detection, high sensitivity, and high selectivity [5]. However, not many food quality test laboratories have this instrumentation.…”

Section: Indonesian Journal Of Chemical Analysismentioning

confidence: 99%

“…One of the contaminations of non-halal materials comes from pork products in meat products and their derivatives. Pork fat can be identified through DNA analysis using polymerase chain reaction (PCR), DNA barcoding, loop-mediated isothermal amplification (LAMP), and biosensor method [5], [6]. The DNA method is reported as capable of accurately detecting pork DNA,…”

Section: Introductionmentioning

confidence: 99%

Identification of Fat in Pork Using Fourier Transform Infrared Spectrum and GC-MS

Yulirohyami,

Maulidatunnisa,

Pusparani

et al. 2023

Ind. J. Chem. Anal

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Halal food safety issues receive serious attention. Methods for analyzing contamination of non-halal ingredients, such as lard, must be developed to assist in the halal authentication process for food. This study was conducted to compare the fat profile of pork thighs using FT-IR and GC-MS. The pork is dried in the oven and then extracted using n-hexane. The resulting fat was characterized by FT-IR. Determination of fatty acids was carried out by GC-MS using a derivatization technique. Based on the results of the study showed that the infrared pattern of lard can be identified from the difference in absorption intensity. Pork fat has a higher unsaturated fatty acid content than beef and chicken. Infrared spectra can confirm the presence of -C=C- bonds at 3005, 1745, 1116, 1550, and 722 cm-1. The FTIR data shows that there are larger unsaturated fatty acid groups in pork fat than in beef and chicken fat. This procedure is a non-destructive method that can be carried out quickly, and cheaply, and can be developed into a routine procedure for halal authentication of food. Based on the results of unsaturated fatty acid testing with GC-MS it can show the content of oleic acid (C18:1) contained in pork. The results of mapping the content of free fatty acids in pork can be used to complete halal authentication data easily and quickly.

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