Sensitivity of polymerase chain reaction (PCR)-southern hybridization and conventional PCR analysis for Halal authentication of gelatin capsules

Sahilah Abd. Mutalib, Nursheila Mustafa Muin, Aminah Abdullah, Osman Hassan, Wan Aida Wan Mustapha, Norrakiah Abdullah Sani, Mohamad Yusof Maskat

Research output: Contribution to journalArticle

22 Citations (Scopus)

Abstract

Halal authentication of gelatin capsules was conducted using polymerase chain reaction (PCR)-southern hybridization on chip and conventional PCR analysis. The primers used in PCR-southern hybridization were targeted on mitochondria DNA (mtDNA) of cytochrome b (cyt b) gene and its amplicon was 276bp. In conventional PCR, three pairs of mtDNA primers targeted cyt b, cytochrome oxidase II (COII) and ATP6 gene were tested, resulting of 398, 212 and 83bp amplicons, respectively. Of 20 brands examined using PCR-southern hybridization, 6 capsules (C1-C6) were found to be porcine DNA positive but none were positive using conventional PCR method. The sensitivity of each primer in the detection of porcine DNA was 0.25ng (cyt b), 0.1ng (COII), 0.001ng (Olipro™ Chip) and 0.0001ng (ATP6). Results demonstrated that the PCR-southern hybridization on chip was useful and reliable for verifying porcine DNA in gelatin capsules compared to conventional PCR.

Original languageEnglish
Pages (from-to)714-719
Number of pages6
JournalLWT - Food Science and Technology
Volume63
Issue number1
DOIs
Publication statusPublished - 1 Sep 2015

Fingerprint

Gelatin
gelatin
Southern blotting
Capsules
polymerase chain reaction
Polymerase Chain Reaction
Cytochromes b
cytochrome b
Swine
DNA
Electron Transport Complex IV
cytochrome-c oxidase
swine
Mitochondria
mitochondria
DNA Primers
DNA primers
Genes
genes

Keywords

  • Conventional PCR
  • Gelatin capsules
  • Halal authentication
  • PCR-southern hybridization
  • Porcine DNA detection

ASJC Scopus subject areas

  • Food Science

Cite this

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title = "Sensitivity of polymerase chain reaction (PCR)-southern hybridization and conventional PCR analysis for Halal authentication of gelatin capsules",
abstract = "Halal authentication of gelatin capsules was conducted using polymerase chain reaction (PCR)-southern hybridization on chip and conventional PCR analysis. The primers used in PCR-southern hybridization were targeted on mitochondria DNA (mtDNA) of cytochrome b (cyt b) gene and its amplicon was 276bp. In conventional PCR, three pairs of mtDNA primers targeted cyt b, cytochrome oxidase II (COII) and ATP6 gene were tested, resulting of 398, 212 and 83bp amplicons, respectively. Of 20 brands examined using PCR-southern hybridization, 6 capsules (C1-C6) were found to be porcine DNA positive but none were positive using conventional PCR method. The sensitivity of each primer in the detection of porcine DNA was 0.25ng (cyt b), 0.1ng (COII), 0.001ng (Olipro™ Chip) and 0.0001ng (ATP6). Results demonstrated that the PCR-southern hybridization on chip was useful and reliable for verifying porcine DNA in gelatin capsules compared to conventional PCR.",
keywords = "Conventional PCR, Gelatin capsules, Halal authentication, PCR-southern hybridization, Porcine DNA detection",
author = "{Abd. Mutalib}, Sahilah and Muin, {Nursheila Mustafa} and Aminah Abdullah and Osman Hassan and {Wan Mustapha}, {Wan Aida} and {Abdullah Sani}, Norrakiah and Maskat, {Mohamad Yusof}",
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T1 - Sensitivity of polymerase chain reaction (PCR)-southern hybridization and conventional PCR analysis for Halal authentication of gelatin capsules

AU - Abd. Mutalib, Sahilah

AU - Muin, Nursheila Mustafa

AU - Abdullah, Aminah

AU - Hassan, Osman

AU - Wan Mustapha, Wan Aida

AU - Abdullah Sani, Norrakiah

AU - Maskat, Mohamad Yusof

PY - 2015/9/1

Y1 - 2015/9/1

N2 - Halal authentication of gelatin capsules was conducted using polymerase chain reaction (PCR)-southern hybridization on chip and conventional PCR analysis. The primers used in PCR-southern hybridization were targeted on mitochondria DNA (mtDNA) of cytochrome b (cyt b) gene and its amplicon was 276bp. In conventional PCR, three pairs of mtDNA primers targeted cyt b, cytochrome oxidase II (COII) and ATP6 gene were tested, resulting of 398, 212 and 83bp amplicons, respectively. Of 20 brands examined using PCR-southern hybridization, 6 capsules (C1-C6) were found to be porcine DNA positive but none were positive using conventional PCR method. The sensitivity of each primer in the detection of porcine DNA was 0.25ng (cyt b), 0.1ng (COII), 0.001ng (Olipro™ Chip) and 0.0001ng (ATP6). Results demonstrated that the PCR-southern hybridization on chip was useful and reliable for verifying porcine DNA in gelatin capsules compared to conventional PCR.

AB - Halal authentication of gelatin capsules was conducted using polymerase chain reaction (PCR)-southern hybridization on chip and conventional PCR analysis. The primers used in PCR-southern hybridization were targeted on mitochondria DNA (mtDNA) of cytochrome b (cyt b) gene and its amplicon was 276bp. In conventional PCR, three pairs of mtDNA primers targeted cyt b, cytochrome oxidase II (COII) and ATP6 gene were tested, resulting of 398, 212 and 83bp amplicons, respectively. Of 20 brands examined using PCR-southern hybridization, 6 capsules (C1-C6) were found to be porcine DNA positive but none were positive using conventional PCR method. The sensitivity of each primer in the detection of porcine DNA was 0.25ng (cyt b), 0.1ng (COII), 0.001ng (Olipro™ Chip) and 0.0001ng (ATP6). Results demonstrated that the PCR-southern hybridization on chip was useful and reliable for verifying porcine DNA in gelatin capsules compared to conventional PCR.

KW - Conventional PCR

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KW - Halal authentication

KW - PCR-southern hybridization

KW - Porcine DNA detection

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