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Identification of Pork Contamination in Meatballs of Indonesia Local Market Using Polymerase Chain Reaction-Restriction Fragment Length Polymorphism (PCR-RFLP) Analysis.

Erwanto Y, Abidin MZ, Sugiyono EY, Rohman A - Asian-australas. J. Anim. Sci. (2014)

Bottom Line: In conclusion, specific PCR amplification of cytochrome b gen and cleaved by BseDI restriction enzymes seems to be a powerful technique for the identification of pork presence in meatball because of its simplicity, specificity and sensitivity.Furthermore, pork contamination intended for commercial products of sausage, nugget, steak and meat burger can be checked.The procedure is also much cheaper than other methods based on PCR, immunodiffusion and other techniques that need expensive equipment.

View Article: PubMed Central - PubMed

Affiliation: Halal Products Research Center, Gadjah Mada University, Yogyakarta 55281, Indonesia .

ABSTRACT
This research applied and evaluated a polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) using cytochrome b gene to detect pork contamination in meatballs from local markets in Surabaya and Yogyakarta regions, Indonesia. To confirm the effectiveness and specificity of this fragment, thirty nine DNA samples from different meatball shops were isolated and amplified, and then the PCR amplicon was digested by BseDI restriction enzyme to detect the presence of pork in meatballs. BseDI restriction enzyme was able to cleave porcine cytochrome b gene into two fragments (131 bp and 228 bp). Testing the meatballs from the local market showed that nine of twenty meatball shops in Yogyakarta region were detected to have pork contamination, but there was no pork contamination in meatball shops in Surabaya region. In conclusion, specific PCR amplification of cytochrome b gen and cleaved by BseDI restriction enzymes seems to be a powerful technique for the identification of pork presence in meatball because of its simplicity, specificity and sensitivity. Furthermore, pork contamination intended for commercial products of sausage, nugget, steak and meat burger can be checked. The procedure is also much cheaper than other methods based on PCR, immunodiffusion and other techniques that need expensive equipment.

No MeSH data available.


Related in: MedlinePlus

Total genomic DNA extracted from local market meatballs. M: marker 100 bp DNA ladder (Invitrogen); A and B: genomic DNA of meatballs from different meatball shops in Yogyakarta (lane 1 to 20); C and D: genomic DNA of meatballs from Surabaya Region (lane 1 to 19); b: Total DNA of raw beef, and p: Total DNA of raw pork.
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f1-ajas-27-10-1487: Total genomic DNA extracted from local market meatballs. M: marker 100 bp DNA ladder (Invitrogen); A and B: genomic DNA of meatballs from different meatball shops in Yogyakarta (lane 1 to 20); C and D: genomic DNA of meatballs from Surabaya Region (lane 1 to 19); b: Total DNA of raw beef, and p: Total DNA of raw pork.

Mentions: Genomic DNA was used as template for amplification by PCR with the universal primer. The first step of the protocol for PCR analysis is the extraction of genomic DNA. The extraction genomic DNA from the commercial meatballs samples was reliable although there were some smearing of the DNA in the samples. Figure 1 showed the electrophoretic patterns of genomic DNA from meatballs of Indonesia local market.


Identification of Pork Contamination in Meatballs of Indonesia Local Market Using Polymerase Chain Reaction-Restriction Fragment Length Polymorphism (PCR-RFLP) Analysis.

Erwanto Y, Abidin MZ, Sugiyono EY, Rohman A - Asian-australas. J. Anim. Sci. (2014)

Total genomic DNA extracted from local market meatballs. M: marker 100 bp DNA ladder (Invitrogen); A and B: genomic DNA of meatballs from different meatball shops in Yogyakarta (lane 1 to 20); C and D: genomic DNA of meatballs from Surabaya Region (lane 1 to 19); b: Total DNA of raw beef, and p: Total DNA of raw pork.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4150182&req=5

f1-ajas-27-10-1487: Total genomic DNA extracted from local market meatballs. M: marker 100 bp DNA ladder (Invitrogen); A and B: genomic DNA of meatballs from different meatball shops in Yogyakarta (lane 1 to 20); C and D: genomic DNA of meatballs from Surabaya Region (lane 1 to 19); b: Total DNA of raw beef, and p: Total DNA of raw pork.
Mentions: Genomic DNA was used as template for amplification by PCR with the universal primer. The first step of the protocol for PCR analysis is the extraction of genomic DNA. The extraction genomic DNA from the commercial meatballs samples was reliable although there were some smearing of the DNA in the samples. Figure 1 showed the electrophoretic patterns of genomic DNA from meatballs of Indonesia local market.

Bottom Line: In conclusion, specific PCR amplification of cytochrome b gen and cleaved by BseDI restriction enzymes seems to be a powerful technique for the identification of pork presence in meatball because of its simplicity, specificity and sensitivity.Furthermore, pork contamination intended for commercial products of sausage, nugget, steak and meat burger can be checked.The procedure is also much cheaper than other methods based on PCR, immunodiffusion and other techniques that need expensive equipment.

View Article: PubMed Central - PubMed

Affiliation: Halal Products Research Center, Gadjah Mada University, Yogyakarta 55281, Indonesia .

ABSTRACT
This research applied and evaluated a polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) using cytochrome b gene to detect pork contamination in meatballs from local markets in Surabaya and Yogyakarta regions, Indonesia. To confirm the effectiveness and specificity of this fragment, thirty nine DNA samples from different meatball shops were isolated and amplified, and then the PCR amplicon was digested by BseDI restriction enzyme to detect the presence of pork in meatballs. BseDI restriction enzyme was able to cleave porcine cytochrome b gene into two fragments (131 bp and 228 bp). Testing the meatballs from the local market showed that nine of twenty meatball shops in Yogyakarta region were detected to have pork contamination, but there was no pork contamination in meatball shops in Surabaya region. In conclusion, specific PCR amplification of cytochrome b gen and cleaved by BseDI restriction enzymes seems to be a powerful technique for the identification of pork presence in meatball because of its simplicity, specificity and sensitivity. Furthermore, pork contamination intended for commercial products of sausage, nugget, steak and meat burger can be checked. The procedure is also much cheaper than other methods based on PCR, immunodiffusion and other techniques that need expensive equipment.

No MeSH data available.


Related in: MedlinePlus