scholarly journals Sus sp. DNA Encoding cyt b Gene Detection Test on Meat Grinding Samples Using Conventional PCR

2020 ◽  
Vol 2 (2) ◽  
pp. 40-45
Author(s):  
Miftahul Lathif Adzakiyyi ◽  
Tri Susilowati ◽  
Saiku Rokhim ◽  
Yuanita Rachmawati
Keyword(s):  
Conventional Polymerase Chain Reaction ◽  
Positive Control ◽  
Pcr Analysis ◽  
Processed Meat ◽  
Cyt B ◽  
Dna Encoding ◽  
Pcr Method ◽  
The Government ◽  
C 30 ◽  
Cyt B Gene

Micro-entrepreneurs with basic ingredients of processed meat such as meatball who do not have a meat grinder, generally using meat grinder at the public market. The problem that occurs is that there is no clear regulation from the Government regarding the guarantee of the halal meat grinding in the Regional Company. This needs to be enhanced as a study, considering that the grinding material does not only come from Halal substances. The purpose of this study was to test pig DNA in meat grinding samples at PD Pasar Surya Surabaya City by using the conventional Polymerase Chain Reaction (PCR) method. DNA was isolated from 11 PD Pasar Surya meat grinding samples, then spectrophotometry was performed. Spectrophotometry results showed that all samples have high DNA concentrations. The primer used is the cyt b pig gene encoder. Predenaturation is performed at a temperature of 95°C-5 minutes, denaturation of 95°C-45 seconds, annealing 60°C-30 seconds, extension 72°C-40 seconds, and post extension 72°C-5 minutes. The results of PCR analysis were determined by the emergence of DNA bands of ± 149 bp as markers of pig DNA. The results showed negative on sample or no pig contamination in 11 samples tested. While the pig sample as positive control showed a band of ± 149 bp. These results prove that at 11 points of the location of meat grinding there is no contamination of pig DNA.

scholarly journals Sus sp. DNA ENCODING cyt b GENE DETECTION TEST ON SOFT GEL CANDY SAMPLES USING PCR METHOD

2021 ◽  
Vol 4 (1) ◽  
pp. 14
Author(s):  
Latifatoel Chilmi ◽  
Tri Susilowati ◽  
Yuanita Rachmawati ◽  
Saiku Rokhim ◽  
Inggrit Tyautari
Keyword(s):  
Aging Treatment ◽  
Positive Control ◽  
Cyt B ◽  
Dna Encoding ◽  
Gene Encoding ◽  
Animal Bone ◽  
Pcr Product ◽  
Pcr Method ◽  
Main Components ◽  
Cyt B Gene

Softgel candy is soft-textured confectionery processed by the addition of several components such as gum, pectin, starch and gelatin, to obtain a supple product and packed after aging treatment first. Gelatin is one of the main components in the manufacture of soft candy derived from the hydrolysis of collagen connective tissue and animal bone that serves as the nature of gelling agents, stabilizers or emulsifiers. However, the gelatin used in products not yet labeled halal Indonesian Council of Ulama (MUI) is particularly vulnerable to pork gelatin, since pork gelatin is cheaper than cattle. The purpose of this study was to test the contaminants of pig DNA on 17 samples of soft candles not labeled halal MUI. This research used Polymerase Chain Reaction (PCR) method. Seventeen samples were isolated by DNA, then spectrophotometry was performed, followed by PCR. The PCR product is run electrophoresis. Visualize the DNA with a UV gel documentation. Primer used is primer gene encoding cyt b DNA pork. Results showed that 17 samples were negative contaminants, while the positive control of pork showed a DNA band of 149 bp. This shows that Softgel Candy 17 samples do not contain pork gelatin.

scholarly journals Detection of Salmonella typhimurium ATCC 14028 in Powder Prepared Traditional Medicines Using Real-Time PCR

2021 ◽  
Vol 4 (3) ◽  
pp. 178-183
Author(s):  
Alfi Sophian ◽  
Ratna Purwaningsih ◽  
Muindar Muindar ◽  
Eka Putri Juniarti Igirisa ◽  
Muhammad Luthfi Amirullah
Keyword(s):  
Salmonella Typhimurium ◽  
Real Time ◽  
Real Time Pcr ◽  
Positive Control ◽  
Negative Control ◽  
Pcr Analysis ◽  
Direct Pcr ◽  
Traditional Medicines ◽  
Pcr Method ◽  
Alternative Testing

The detection of Salmonella typhimurium ATCC 14028 using real-time PCR on powdered traditional medicinal products was carried out in the microbiology and molecular biology testing laboratory of the Food and Drug Administration in Gorontalo. This research aims to provide a reference for alternative testing methods in testing the products of traditional powder preparations on the market. The sample consisted of 10 traditional powder preparations spiked with positive control of S. typhimurium ATCC 14028 phase 2. The method used in the study was real-time PCR analysis using the SYBR® Green method, while DNA isolation using the direct PCR method. Data analysis was performed by analyzing the sample's melting temperature (Tm) curve and comparing it with positive control. The results showed that S. typhimurium ATCC 14028 was detected in samples at an average Tm value of 84.18°C, with ranges of 84.0-84.5°C. For positive control, the Tm value was at 85.2°C, while for the negative control, the Tm value was not detected. Based on these data, it can be concluded that S. typhimurium ATCC 14028 in traditional medicine products powder preparations can be detected using real-time PCR.

scholarly journals AUTENTIKASI DAGING AYAM SEGAR DARI KONTAMINASI DAGING BABI MENGGUNAKAN GEN CYT-B DENGAN ANALISIS DUPLEX- POLYMERASE CHAIN REACTION

2017 ◽  
Vol 41 (2) ◽  
pp. 113
Author(s):  
Bayu Setya Hertanto ◽  
Rizky Aulia Fitra ◽  
Lilik Retna Kartikasari ◽  
Muhammad Cahyadi
Keyword(s):  
Meat Products ◽  
Chicken Meat ◽  
Pcr Analysis ◽  
Processed Meat ◽  
Duplex Pcr ◽  
Cyt B ◽  
Polymerase Chain ◽  
Contamination Levels ◽  
The City ◽  
Dna Isolation Protocol

Halal is one of important aspects in consumer protection. Meat and processed meat products are food that should be controlled strictly because those are prone to be adulterated by pork contamination. Therefore, it is necessary to provide detection technique which is accurate, fast and cheap. The objective of this research was to identify the presence of impurities of pork meat on raw chicken meat using gene Cyt-b with duplex-PCR analysis. This research used six samples of raw chicken meat and raw pork. Raw chicken meat was bought from supermarkets in the city of Surakarta and raw pork was obtained from pig slaughterhouse. The percentage of raw pork contamination on raw chicken meat was designed as much as 1, 5, 10, and 25%, respectively. The DNA genome was isolated according to DNA isolation protocol from Genomic DNA Mini Kit. In addition, duplex-PCR was performed based on protocol of KAPA2G Fast Multiplex PCR kit. The data was descriptively analyzed by directly looking the DNA bands on the gel documentation apparatus. The result showed that specific DNA bands for chicken and pig were completely appeared on 1.5% of agarose gels. Duplex-PCR detect contamination of pork on raw meat of chicken at all contamination levels. This research proved that the duplex-PCR detect the contamination of pork until the level of 1%.

scholarly journals Multiplex PCR Method of Detecting Pork to Guarantee Halal Status in Meat Processed Products

2019 ◽  
Vol 7 (3) ◽  
pp. 96-101
Author(s):  
M. Indriati ◽  
E. Yuniarsih
Keyword(s):  
Multiplex Pcr ◽  
Meat Product ◽  
Dna Amplification ◽  
Processed Meat ◽  
Target Sequence ◽  
Cyt B ◽  
Traditional Markets ◽  
Pcr Multiplex ◽  
Processed Products ◽  
Cyt B Gene

One of the halal parameters of food is must be free from pork among from the basic ingredients, additives and the manufacturing process. The aim of this study was ensure the halal status in the form of presence or absence of pork in processed meat product (meatballs and sausages) in traditional markets in Pandeglang Regency. PCR multiplex was a PCR technique that used several primers together in one reaction to amplify several target sequence. The genes often used as markers of animal or meat types include cytochrome b (cyt b), the existence of sequence variations in cyt b causes these genes are widely used as markers to distinguish material from different types of animals. The results showed that the cyt b gene proved successful in amplified DNA from beef and pork with different fragment lengths in the DNA mix of the 2 types of livestock in one reaction so that 2 DNA bands formed with different lengths according to the length of the fragment from each animal. From the results of research showed beef and pork control there was two fragments such as 274 bp for beef and 389 bp for pork. Multiplex PCR testing on meatball samples showed that the meatballs were tested 100% positive containing beef and 0% containing pork DNA. While testing on sausage products, there were one sausage brand that showed the results of DNA amplification of 398pb, which means the product was positive containing pork.

scholarly journals Amino acid diversity on the basis of cytochrome b gene in Kacang and Ettawa Grade goats

2017 ◽  
Vol 42 (3) ◽  
pp. 135 ◽  
Author(s):  
D. A. Lestari ◽  
S. Sutopo ◽  
E. Kurnianto
Keyword(s):  
Amino Acids ◽  
Amino Acid ◽  
Genetic Marker ◽  
Cytochrome B ◽  
Genomic Dna ◽  
Cyt B ◽  
Specific Amino Acid ◽  
Pcr Method ◽  
Cyt B Gene ◽  
Amino Acid Diversity

The objectives of study were to identify and assess the amino acid diversity of Cytochrome b (Cyt b) gene, genetic marker and characteristic of specific amino acid in Kacang and Ettawa Grade goat. Nineteen heads of Kacang goat (KG) and twelve heads of Ettawa Grade goat (EG) were purposively sampled. The genomic DNA was isolated by Genomic DNA Mini Kit (Geneaid) and amplified Cyt b using PCR method with CytbCapF and CytbCapR primers and was sequenced. The results showed that there were two specific amino acids that distinguish KG and EG goat with C. hircus and C. aegagrus and four specific amino acids that distinguish KG and EG goat with C. falconeri, but there were no specific amino acids can be used as a genetic marker to distinguish between Kacang and EG goat. In conclusion, specific amino acids in Cyt b gene can be used as a genetic marker among KG and EG goat with 3 goat others comparator.

scholarly journals Detection of DNA pork in processed meat products with real-time polymerase chain reaction

Food Research ◽  
2020 ◽  
Vol 4 (5) ◽  
pp. 1719-1725
Author(s):  
W.P. Rahayu ◽  
A.R.W. Dianti ◽  
Nurjanah S. ◽  
N. Pusparini ◽  
W. Adhi
Keyword(s):  
Polymerase Chain Reaction ◽  
Real Time ◽  
Melting Curve ◽  
Meat Products ◽  
Processed Meat ◽  
Cyt B ◽  
Chain Reaction ◽  
Ct Value ◽  
Polymerase Chain ◽  
Cyt B Gene

Real-time polymerase chain reaction (qPCR) technique is a suitable method for identifying animal species in processed meat because of its ability to amplify a few fragments of DNA. A specific fragment of pork (mitochondrial cytochrome b gene (cytb)) was used as a DNA ladder. This study aimed to evaluate the use of a cyt-b gene generated primer for detecting the presence of pork in processed meat products by qPCR and determining the threshold cycle cut-off. The evaluation of the primer effectiveness was performed by threshold cycle (Ct) value, amplicon size by electrophoresis and melting curve. Two corned (A, B) and two jerkies (C, D) collected from the market were used as the sample. Genomic DNA from samples, fresh beef (as negative control) and fresh pork (as positive control) were extracted using Qiagen Kits. Amplification condition for 50 cycles of the cyt-b gene was performed as the initial step at 95°C for 10 mins, denaturation step at 95°C for 15 s, annealing step at 55°C for 60 s, extension step at 72°C for 30 s and post-PCR at 72°C for 3 mins. The threshold cycle (Ct) cut-off less than 30 confirmed as pork positive. The result obtained indicated that sample A and D were pork negative, with Ct value respectively 40.73 and 43.59. Melting temperatures of amplicon were ranged from 79.5 to 80.5°C, only differed by 1°C, and the amplicon electrophoresis resulting in a single band of the same size (149 bp). Hence, the melting curve analysis and electrophoresis of PCR products were not able to differentiate between pork and beef.

PCR Detection of Alternaria spp. in Processed Foods, Based on the Internal Transcribed Spacer Genetic Marker

2011 ◽  
Vol 74 (2) ◽  
pp. 240-247 ◽  
Author(s):  
MIGUELÁNGEL PAVÓN ◽  
ISABEL GONZÁLEZ ◽  
MARÍA ROJAS ◽  
NICOLETTE PEGELS ◽  
ROSARIO MARTÍN ◽  
...  
Keyword(s):  
Food Processing ◽  
Internal Transcribed Spacer ◽  
Rapid Identification ◽  
Food Samples ◽  
Pcr Analysis ◽  
Infant Food ◽  
Rrna Gene ◽  
Tomato Pulp ◽  
Pcr Method ◽  
Fungal Contaminants

The genus Alternaria is considered one of the most important fungal contaminants of vegetables, fruits, and cereals, producing several mycotoxins that can withstand food processing methods. Conventional methods for Alternaria identification and enumeration are laborious and time-consuming, and they might not detect toxigenic molds inactivated by food processing. In this study, a PCR method has been developed for the rapid identification of Alternaria spp. DNA in foodstuffs, based on oligonucleotide primers targeting the internal transcribed spacer (ITS) 1 and ITS2 regions of the rRNA gene. The specificity of the Alternaria-specific primer pair designed (Dir1ITSAlt–Inv1ITSAlt) was verified by PCR analysis of DNA from various Alternaria spp., and also from several fungal, bacterial, yeast, animal, and plant species. The detection limit of the method was 102 CFU/ml in viable culture, heated culture, or experimentally inoculated tomato pulp. The applicability of the method for detection of Alternaria spp. DNA in foodstuffs was assessed by testing several commercial samples. Alternaria DNA was detected in 100% of spoiled tomato samples, 8% of tomato products, and 36.4% of cereal-based infant food samples analyzed.

scholarly journals Induction of mucosal and systemic immune response by single-dose oral immunization with biodegradable microparticles containing DNA encoding HBsAg

2005 ◽  
Vol 86 (3) ◽  
pp. 601-610 ◽  
Author(s):  
Xiao-Wen He ◽  
Fang Wang ◽  
Lei Jiang ◽  
Jun Li ◽  
Shan-kui Liu ◽  
...  
Keyword(s):  
Single Dose ◽  
Plasmid Dna ◽  
Dna Vaccines ◽  
Oral Immunization ◽  
Laser Scanning Microscopy ◽  
Confocal Laser ◽  
Pcr Analysis ◽  
Target Antigen ◽  
Dna Encoding ◽  
Plga Microparticles

The purpose of this work was to assess the ability of plasmid DNA encoding hepatitis B virus (HBV) HBsAg encapsulated in poly(dl-lactide-co-glycolic acid) (PLGA) microparticles to induce local and systemic HBsAg-specific immunity following a single dose of oral immunization. RT-PCR analysis demonstrated prolonged transcription of plasmid DNA, consistent with the sustained expression and presentation of target antigen observed by confocal laser scanning microscopy, in gut-associated lymphocyte tissue (GALT) from mice immunized orally with plasmid DNA encapsulated into PLGA microparticles. Oral administration of PLGA-DNA microparticles induced a long-lasting and stable antigen-specific antibody response, both serum total antibody and intestinal IgA, in BALB/c mice. Mice immunized orally exhibited antigen-specific gamma interferon production and cytotoxic T lymphocyte responses in spleen and GALT after restimulation in vitro with HBsAg or tumour cells stably expressing HBsAg. In contrast, naked DNA vaccines given by intramuscular injection induced only systemic cellular and humoral responses to HBsAg, which were much lower than the responses elicited by oral DNA encapsulated in PLGA microparticles at equivalent doses. The results are encouraging with regard to obtaining good compliance and vaccination coverage with candidate plasmid DNA vaccines, especially in developing countries.

scholarly journals Genetic characterization and phylogenetic study of Indonesian indigenous catfish based on mitochondrial cytochrome B gene

2020 ◽  
Vol 13 (1) ◽  
pp. 96-103
Author(s):  
Dorothea Vera Megarani ◽  
Herjuno Ari Nugroho ◽  
Zahrah Prawita Andarini ◽  
Yura Dwi Risa B. R. Surbakti ◽  
Rini Widayanti
Keyword(s):  
Amino Acids ◽  
Amino Acid ◽  
Cytochrome B ◽  
Genetic Characterization ◽  
Phylogenetic Study ◽  
Cyt B ◽  
Phylogenetic Structure ◽  
Mitochondrial Cytochrome B ◽  
Sperata Seenghala ◽  
Cyt B Gene

Aim: This study aimed to determine the genetic characterization and phylogenetic structure of Indonesian indigenous catfish using cytochrome B (Cyt B) sequences. Materials and Methods: The genomes of 26 catfishes caught from nine rivers from nine different geographical locations around Indonesia were analyzed. The tissue isolation method was used to isolate the total genome of the fishes. Furthermore, polymerase chain reaction was done to amplify the mtDNA Cyt B using the CytBF and CytBR primers. Following sequencing, the analysis of genetic variation and the phylogenetic relationship was performed using MEGA version X software. Results: Cyt B gene sequencing attained a total of 1139 nucleotides encrypting 379 amino acids for all samples. The ClustalW alignment program using MEGA X software revealed 395 substituted nucleotides, which then translated into 63 amino acid variation sites among all 26 samples. No amino acids in catfish BB were different compared to catfish PM, MP, and KR2,3. Catfish MS had one modified amino acid; KR1 and KS had two different amino acids; BF had 38 different amino acids; EM had 31 different amino acids; and BSBJ had 26 different amino acids compared to catfish BB. The most significant alteration of amino acids was between catfish EM and BF (49 amino acids). Conclusion: Indonesian catfish were divided into five clades based on the Cyt B gene. Samples KR and MP (Sumatra); MS and BB (Kalimantan); and PM (Java) were clustered with Hemibagrus nemurus and Hemibagrus wyckioides (Bagridae family). Samples from Kalimantan (KS) and one sample of KR (KR1) from Sumatra were clustered with Sperata seenghala and Hemibagrus spilopterus (Bagridae family). Samples from Java (BSBJ) were clustered with Pseudolais pleurotaenia (Pangasiidae family). Samples EM (Java) were together with Mystus cavasius (Bagridae family). Samples from West Papua were clustered with Potamosilurus latirostris (Ariidae family).

scholarly journals Kajian Penanda Genetik Gen Sitokrom b DNA Mitokondria Ikan Lais dari Sungai Kampar Riau

2018 ◽  
Vol 10 (1) ◽  
pp. 6
Author(s):  
Roza Elvyra ◽  
Dedy Duryadi Solihin
Keyword(s):  
Species Diversity ◽  
Molecular Marker ◽  
Phylogenetic Relationship ◽  
Cytochrome B ◽  
Mitochondrial Cytochrome ◽  
Cyt B ◽  
Phylogenetic Marker ◽  
Mitochondrial Cytochrome B ◽  
Cyt B Gene

The mitochondrial cytochrome b (cyt-b) gene as a phylogenetic marker of lais fish Kryptopterus schilbeides from Kampar River in Riau has been studied. This is a prelimininary research on the utility of cyt-b gene as a molecular marker to obtain species diversity and phylogenetic relationship among Kryptopterus fishes from Kampar River. The primers of L14841 and H15149 were used to amplify the cyt-b gene. The results showed that K. schilbeides has isoleusine at site-81 and metionine at site-114; K. schilbeides from Kampar River and K. schilbeides from GenBank form a phylogeny cluster at 45% value.

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