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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

PCR products of cytochrome b gene fragments 359 bp long in samples from different meatballs products separated by 2% high-resolution agarose gel electrophoresis, M: marker 100 bp DNA ladder (Invitrogen), A–B: PCR products of cytochrome b gene from twenty of meatball samples from Yogyakarta region (1–20), C–D: PCR products of cyt b gene from nineteen of meatball samples from Surabaya region (1–19), b: PCR product of raw beef DNA, and p: PCR product of raw pork DNA. PCR, polymerase chain reaction.
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f2-ajas-27-10-1487: PCR products of cytochrome b gene fragments 359 bp long in samples from different meatballs products separated by 2% high-resolution agarose gel electrophoresis, M: marker 100 bp DNA ladder (Invitrogen), A–B: PCR products of cytochrome b gene from twenty of meatball samples from Yogyakarta region (1–20), C–D: PCR products of cyt b gene from nineteen of meatball samples from Surabaya region (1–19), b: PCR product of raw beef DNA, and p: PCR product of raw pork DNA. PCR, polymerase chain reaction.

Mentions: The PCR has the potential sensitivity and specificity to attain detection of a target sequence from template DNA. The PCR products of the cytochrome b gene as an amplicon from the meatball DNA from thirty nine locations of meatball shops are shown in Figure 2. The cytochrome b gene amplified by PCR from a meatball after boiling resulted in a DNA fragment of approximately 360 bp. These small amplicons are ideal for use with processed foods where DNA is commonly degraded. This result indicated that both pig and beef DNA in processed meat was successfully amplified in the PCR reaction.


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)

PCR products of cytochrome b gene fragments 359 bp long in samples from different meatballs products separated by 2% high-resolution agarose gel electrophoresis, M: marker 100 bp DNA ladder (Invitrogen), A–B: PCR products of cytochrome b gene from twenty of meatball samples from Yogyakarta region (1–20), C–D: PCR products of cyt b gene from nineteen of meatball samples from Surabaya region (1–19), b: PCR product of raw beef DNA, and p: PCR product of raw pork DNA. PCR, polymerase chain reaction.
© Copyright Policy
Related In: Results  -  Collection

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

f2-ajas-27-10-1487: PCR products of cytochrome b gene fragments 359 bp long in samples from different meatballs products separated by 2% high-resolution agarose gel electrophoresis, M: marker 100 bp DNA ladder (Invitrogen), A–B: PCR products of cytochrome b gene from twenty of meatball samples from Yogyakarta region (1–20), C–D: PCR products of cyt b gene from nineteen of meatball samples from Surabaya region (1–19), b: PCR product of raw beef DNA, and p: PCR product of raw pork DNA. PCR, polymerase chain reaction.
Mentions: The PCR has the potential sensitivity and specificity to attain detection of a target sequence from template DNA. The PCR products of the cytochrome b gene as an amplicon from the meatball DNA from thirty nine locations of meatball shops are shown in Figure 2. The cytochrome b gene amplified by PCR from a meatball after boiling resulted in a DNA fragment of approximately 360 bp. These small amplicons are ideal for use with processed foods where DNA is commonly degraded. This result indicated that both pig and beef DNA in processed meat was successfully amplified in the PCR reaction.

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