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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

Electrophoretic patterns of cytochrome b gene after digestion by BseDI restriction enzymes. M: marker 100 bp DNA ladder, lane 1 to 20: DNA fragment of different meatball samples from twenty meatball shops in Yogyakarta region, b: DNA fragment after digestion of PCR product of raw beef cytochrome b gene, and p: DNA fragment after digestion of PCR product of raw pork cytochrome b gene. PCR, polymerase chain reaction.
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f3-ajas-27-10-1487: Electrophoretic patterns of cytochrome b gene after digestion by BseDI restriction enzymes. M: marker 100 bp DNA ladder, lane 1 to 20: DNA fragment of different meatball samples from twenty meatball shops in Yogyakarta region, b: DNA fragment after digestion of PCR product of raw beef cytochrome b gene, and p: DNA fragment after digestion of PCR product of raw pork cytochrome b gene. PCR, polymerase chain reaction.

Mentions: The PCR product was digested using BseDI restriction enzyme at 55°C for 3 h. The result of RFLP analysis of meatball samples from various shops in Yogyakarta region (Figure 3) indicated some positive pork contamination. Erwanto et al. (2011) reported that cyt b gene from pig species could be digested by BseDI restriction enzyme and 0.1% contamination level of pork could be detected in sausage products using this method. Data indicated pork contamination from nine meatball shops in Yogyakarta region. The contamination by pork was not found in meatballs from various meatball shops in Surabaya region (Figure 4). BseDI cleavage bands visualized in the gel were suitable for the discrimination of pork in processed food. BseDI end nuclease cleaved the cytochrome b gene products of pig species into two DNA fragments of 228 and 131 bp (Figure 3; lane 1, 2, 3, 4, 5, 8, 9, 10, and l8).


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)

Electrophoretic patterns of cytochrome b gene after digestion by BseDI restriction enzymes. M: marker 100 bp DNA ladder, lane 1 to 20: DNA fragment of different meatball samples from twenty meatball shops in Yogyakarta region, b: DNA fragment after digestion of PCR product of raw beef cytochrome b gene, and p: DNA fragment after digestion of PCR product of raw pork cytochrome b gene. PCR, polymerase chain reaction.
© Copyright Policy
Related In: Results  -  Collection

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

f3-ajas-27-10-1487: Electrophoretic patterns of cytochrome b gene after digestion by BseDI restriction enzymes. M: marker 100 bp DNA ladder, lane 1 to 20: DNA fragment of different meatball samples from twenty meatball shops in Yogyakarta region, b: DNA fragment after digestion of PCR product of raw beef cytochrome b gene, and p: DNA fragment after digestion of PCR product of raw pork cytochrome b gene. PCR, polymerase chain reaction.
Mentions: The PCR product was digested using BseDI restriction enzyme at 55°C for 3 h. The result of RFLP analysis of meatball samples from various shops in Yogyakarta region (Figure 3) indicated some positive pork contamination. Erwanto et al. (2011) reported that cyt b gene from pig species could be digested by BseDI restriction enzyme and 0.1% contamination level of pork could be detected in sausage products using this method. Data indicated pork contamination from nine meatball shops in Yogyakarta region. The contamination by pork was not found in meatballs from various meatball shops in Surabaya region (Figure 4). BseDI cleavage bands visualized in the gel were suitable for the discrimination of pork in processed food. BseDI end nuclease cleaved the cytochrome b gene products of pig species into two DNA fragments of 228 and 131 bp (Figure 3; lane 1, 2, 3, 4, 5, 8, 9, 10, and l8).

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