scholarly journals A Novel Multiplex-PCR Assay to Detect Three Non-Halal Meats Contained in Meatball using Mitochondrial 12S rRNA Gene

2020 ◽  
Vol 40 (4) ◽  
pp. 628-635
Author(s):  
Muhammad Cahyadi ◽  
Tommy Wibowo ◽  
Ahmad Pramono ◽  
Zakaria Husein Abdurrahman
2019 ◽  
Vol 44 (1) ◽  
pp. 10 ◽  
Author(s):  
M. Cahyadi ◽  
I. M. Taufik ◽  
A. Pramono ◽  
Z. H. Abdurrahman

The 12S rRNA gene is one of unique regions in mitochondrial genome usually used for phylogenetic studies and species identification. The objective of present study was to develop species specific primers from mitochondrial 12S rRNA gene for identification of dog and rat in beef by using multiplex PCR assay. Three primer pairs of mitochondrial 12S rRNA gene specific for bovine, dog and rat were designed and selected to evaluate their specificity and fidelity. Moreover, a total of twelve DNA samples extracted from meat tissue were also prepared to test those primers using simplex and multiplex PCR. The PCR products were then visualized using 2% of agarose gel under the UV light and three of them were sequenced. In addition, sequence data were analyzed using Clustal Omega software and BLAST. The result showed that simplex PCR assay successfully amplified DNA targets which are respectively indicated by 155 bp (bovine), 244 bp (dog), and 491 bp (rat) of DNA bands. Furthermore, DNA sample sequences were identically similar to reference sequence used in this study. Multiplex and simplex PCR analyses also indicated that these primer pairs specifically amplified DNA target for each species in the samples containing various species. The results suggested that designed primers in this study could be used to identify dog and rat in raw beef containing these species meat. Further experiment should be conducted using meat-processed products and commercial meat products as samples.


2002 ◽  
Vol 22 (2) ◽  
pp. 174-183 ◽  
Author(s):  
Antoinette C. van der Kuyl ◽  
Donato L. Ph. Ballasina ◽  
John T. Dekker ◽  
Jolanda Maas ◽  
Ronald E. Willemsen ◽  
...  

2021 ◽  
Vol 322 ◽  
pp. 01028
Author(s):  
Nao Fukunaga ◽  
Moe Shimizu ◽  
Shinnosuke Teruya ◽  
Nazifa Naziha Razali ◽  
Satoko Nakashima ◽  
...  

DNA barcoding is an effective and powerful tool for taxonomic identification and thus very useful for biodiversity monitoring. This study investigated the usefulness of the mitochondrial 12S-rRNA gene for the DNA barcoding of shelled marine gastropods. To do so, we determined partial 12S-rRNA sequences of 75 vouchered museum specimens from 69 species of shelled gastropods from Japan. The specimens have been identified morphologically, and natural history data catalog. Sequence analyses through BLAST searches, maximum likelihood phylogenetic analysis, and species delimitation analysis suggested that the 12S-rRNA gene is helpful for barcoding shelled marine gastropods. They thus could be helpful to complement barcoding studies using other markers such as COI. The analyses successfully confirmed all samples’ identity at higher taxonomy (subfamily and above), but much less so at the species level. Our result thus also underlines the lingering problem of DNA barcoding: The lack of comprehensive reference databases of sequences. However, since we provided sequences of properly curated, vouchered museum specimens in this study, our result reported here has thus also helped to give taxonomically reliable reference sequences for biodiversity monitoring and identifications of shelled gastropods which include many important fisheries species.


2002 ◽  
Vol 65 (5) ◽  
pp. 780-785 ◽  
Author(s):  
IRENE V. WESLEY ◽  
KAREN M. HARMON ◽  
JAMES S. DICKSON ◽  
ANN RAMOS SCHWARTZ

A multiplex polymerase chain reaction was developed to simultaneously identify Listeria monocytogenes and species of the genus Listeria. Two sets of primers were used, with the first amplifying a 938-bp region of the 16S rRNA gene that is highly conserved in all Listeria species and the second amplifying a 174-bp region of the listeriolysin (hlyA) gene of L. monocytogenes. Thus, isolates of Listeria spp. yield a single 938-bp product, whereas L. monocytogenes isolates yield both the 938-bp product and a 174-bp product. The specificity of the assay was verified with all six Listeria species and 11 serotypes of L. monocytogenes, as well as nonrelated bacteria. The multiplex PCR assay was used to determine the incidence of Listeria spp., especially L. monocytogenes, in mechanically separated turkey samples (n = 150 samples). L. monocytogenes strains were selected by using the University of Vermont two-step enrichment protocol and plating to selective Palcam agar. The multiplex PCR assay was used for verification of presumptive Listeria colonies. Approximately 38% of mechanically separated turkey samples (57 of 150) yielded L. monocytogenes; an additional 18% of these samples (27 of 150) harbored other Listeria spp. Fifty-one percent (29 of 57) of the L. monocytogenes isolates were of serogroup 1, 44% (25 of 57) were of serogroup 4, and 2% (1 of 57) were assigned to serogroups other than 1 and 4.


2010 ◽  
Vol 30 (6) ◽  
pp. 405-411 ◽  
Author(s):  
Emna Mkaouar-Rebai ◽  
Nourhene Fendri-Kriaa ◽  
Nacim Louhichi ◽  
Abdelaziz Tlili ◽  
Chahnez Triki ◽  
...  

Sensorineural hearing loss has been described in association with different mitochondrial multisystemic syndromes, often characterized by an important neuromuscular involvement. Until now, mutations in mitochondrial DNA, especially in the 12S rRNA, the tRNASer(UCN) and the tRNALeu(UUR) genes, were implicated in syndromic or non-syndromic hearing loss either as a primary cause or as predisposing factors. In the present study, we performed a whole mitochondrial genome screening in two unrelated Tunisian families with inherited hearing loss. Results showed the presence of a novel mutation in the mitochondrial 12S rRNA gene in the two probands of these two families who belong to two different haplogroups: L3 and H6a1. The m.735A>G mutation affects a conserved nucleotide of the mitochondrial 12S rRNA gene in primates and other species and had a conservation index of 78.5% (11/14). We also detected known polymorphisms and sic novel mitochondrial variants. The present study confirmed that the mitochondrial 12S rRNA gene is a hot spot for mutations associated with hearing impairment.


2003 ◽  
Vol 1239 ◽  
pp. 75-78
Author(s):  
C Albarrán ◽  
P Martı́n ◽  
O Garcı́a ◽  
P Garcı́a ◽  
L Fernández de Simón ◽  
...  

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