scholarly journals PENGGUNAAN METODE PCR – RFLP (POLYMERASE CHAIN REACTION – RESTRICTION FRAGMENT LENGTH POLYMORFISM) DALAM MENDETEKSI JAMUR DERMATOFIT

2021 ◽  
Vol 14 (1) ◽  
pp. 96-102
Author(s):  
Awaluddin Awaluddin ◽  
Rizalinda Sjahrir ◽  
Farida Ilyas
Keyword(s):  
Polymerase Chain Reaction ◽  
Restriction Fragment ◽  
Fragment Length ◽  
Trichophyton Rubrum ◽  
Chain Reaction ◽  
Pcr Rflp ◽  
Polymerase Chain ◽  
Restriction Fragment Length

ABSTRAK Dermatofitosis adalah salah satu jamur yang terdiri dari tiga genus : Epidermophyton, Trichophyton dan Microsporum. Penelitian ini bertujuan untuk mengidentifikasi jenis jamur dermatofit dengan metode PCR-RFLP. Penelitian ini merupakan penelitian observasional laboratorium dengan dengan menguji 23 sampel yang diperoleh dari beberapa klinik dan sekolah dasar di Makassar. Hasil penelitian menunjukkan bahwa semua sampel teridentifikasi yakni M. canis 26,1%, Trichophyton rubrum 13,1%,T. mentagrophytes 21,6%, T. tonsurans 8,7%, T. verrucosum 4,2% dan spesies unclassified 26,1%. Kami menyarankan Teknik PCR-RFLP dapat digunakan untuk konfirmasi jenis jamur dermatofit.   Kata Kunci : Dermatofitosis, Jamur Dermatofit, PCR – RFLP

scholarly journals Análise de polimorfismo do gene da somatotropina em vacas Nelore e seu efeito sobre o peso à desmama de suas progênies

1999 ◽  
Vol 51 (6) ◽  
pp. 565-570
Author(s):  
F.J.C. Faria ◽  
S.E.F. Guimarães ◽  
R.M.G. Lima ◽  
G.B. Mourão ◽  
L.E.L. Pinheiro
Keyword(s):  
Polymerase Chain Reaction ◽  
Restriction Fragment Length Polymorphism ◽  
Restriction Fragment ◽  
Length Polymorphism ◽  
Fragment Length ◽  
Fragment Length Polymorphism ◽  
Chain Reaction ◽  
Pcr Rflp ◽  
Polymerase Chain ◽  
Restriction Fragment Length

Informações sobre peso à desmama de um rebanho Nelore foram utilizadas após ajuste para idade padrão de 205 dias, sexo da cria, idade da mãe, touro e mês de desmama, para separar as reprodutrizes em dois grupos, cujos filhos diferiam nesse peso. As médias ajustadas pelo método dos quadrados mínimos foram para os grupos pesado (P) e leve (L) de 163,21± 2,18kg e 134,44± 2,18kg, respectivamente, com 41 animais em cada grupo. Essas reprodutrizes foram submetidas à coleta de sangue para estudo de polimorfismos do gene da somatotropina bovina, pela técnica de PCR-RFLP (polymerase chain reaction-restriction fragment length polymorphism). A amplificação de uma região entre o éxon III e V do gene da somatotropina permitiu analisar dois sítios de restrição. Para o sítio do éxon V, todos os animais foram identificados como monomórficos (Leu-Leu). Quanto ao sítio do íntron 3, foi possível identificar os seguintes genótipos 21 (+/-) e 60 (-/-), com as freqüências de 0,13 e 0,87 para os alelos (+) e (-), respectivamente. O peso dos filhos dos animais com o genótipo +/- foi de 152,42± 4,41kg e os -/- 147,60± 2,61kg. Os grupos P e L não diferiram entre si quanto às freqüências alélicas apresentadas. O genótipo das reprodutrizes não afetou o peso à desmama de suas crias, existindo portanto outros efeitos genéticos e não genéticos de maior magnitude.

scholarly journals Polymerase Chain Reaction-Restriction Fragment Length Polymorphism Authentication of Raw Meats from Game Birds

2008 ◽  
Vol 91 (6) ◽  
pp. 1416-1422 ◽  
Author(s):  
María Rojas ◽  
Isabel González ◽  
Violeta Fajardo ◽  
Irene Martín ◽  
Pablo E Hernández ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Restriction Fragment Length Polymorphism ◽  
Restriction Fragment ◽  
Length Polymorphism ◽  
Fragment Length ◽  
Fragment Length Polymorphism ◽  
Chain Reaction ◽  
Pcr Rflp ◽  
Polymerase Chain ◽  
Restriction Fragment Length

Abstract Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis has been applied to the identification of meats from quail (Coturnix coturnix), pheasant (Phasianus colchicus), red-legged partridge (Alectoris rufa), guinea fowl (Numida meleagris), capercaillie (Tetrao urogallus), Eurasian woodcock (Scolopax rusticola), woodpigeon (Columba palumbus), and song thrush (Turdus philomelos). PCR amplification was performed using a set of primers flanking a conserved region of approximately 720 base pairs (bp) from the mitochondrial 12S rRNA gene. Restriction site analysis based on sequence data from this DNA fragment permitted the selection of Alul and BfaI endonucleases for species identification. The restriction profiles obtained when amplicons were digested with the chosen enzymes allowed the unequivocal identification of all game bird species analyzed. However, the use of the PCR-RFLP technique described is limited to raw meat authentication. It is not suitable for cooked products because thermal treatment strongly accelerates DNA degradation leading to difficulties in amplifying the 720 bp fragment.

Polymerase Chain Reaction–Restriction Fragment Length Polymorphism Analysis: A Simple Method for Species Identification in Food

1995 ◽  
Vol 78 (6) ◽  
pp. 1542-1551 ◽  
Author(s):  
Rolf Meyer ◽  
Christiane Höfelein ◽  
Jürg Lüthy ◽  
Urs Candrian
Keyword(s):  
Polymerase Chain Reaction ◽  
Species Identification ◽  
Restriction Fragment ◽  
Fragment Length ◽  
Chain Reaction ◽  
Heat Treated ◽  
Mitochondrial Cytochrome B ◽  
Pcr Rflp ◽  
Polymerase Chain ◽  
Restriction Fragment Length

Abstract The polymerase chain reaction (PCR) technique was applied to meat species identification in marinated and heat-treated or fermented products and to the differentiation of closely related species. DNA was isolated from meat samples by using a DNA-binding resin and was subjected to PCR analysis. Primers used were complementary to conserved areas of the vertebrate mitochondrial cytochrome b (cytb) gene and yielded a 359 base-pair (bp) fragment, including a variable 307 bp region. Restriction endonuclease analysis based on sequence data of those fragments was used for diffferentiation among species. Restriction fragment length polymorphisms (RFLPs) were detected when pig, cattle, wild boar, buffalo, sheep, goat, horse, chicken, and turkey amplicons were cut with Alul, Rsal, Taql, and Hinfl. Analysis of sausages indicates the applicability of this approach to food products containing meat from 3 different species. The PCR–RFLP analytical method detected pork in heated meat mixtures with beef at levels below 1%, and the method was confirmed with porcine- and bovine-specific PCR assays by amplifying fragments of their growth hormone genes. Inter- and intraspecific differences of more than 22 animal species with nearly unknown cytb DNA sequences, including hoofed mammals (ungulates), and poultry were determined with PCR–RFLP typing by using 20 different endonucleases. This typing method allowed the discrimination of game meats, including stag, roe deer, chamois, moose, reindeer, kangaroo, springbok, and other antelopes in marinated and heat-treated products.

Polymerase chain reaction - restriction fragment length polymorphisms for assessing and increasing biodiversity ofFrankiaculture collections

1999 ◽  
Vol 77 (9) ◽  
pp. 1261-1269 ◽  
Author(s):  
Erica Lumini ◽  
Marco Bosco
Keyword(s):  
Polymerase Chain Reaction ◽  
Restriction Fragment ◽  
Fragment Length ◽  
Intergenic Spacer ◽  
Chain Reaction ◽  
Culture Collections ◽  
Length Polymorphisms ◽  
Pcr Rflp ◽  
Polymerase Chain ◽  
Restriction Fragment Length

During the last few years, some Frankia culture collections that maintained a large number of unidentified and uncharacterized Frankia strains were closed because of funding shortages. To reduce the costs of maintenance, we evaluated the biodiversity of half of the Frankia strains from our collection, by polymerase chain reaction - restriction fragment length polymorphisms (PCR-RFLPs) of nifD-nifK intergenic spacer and 16S-23S rDNA intergenic spacer regions. In this way we were able to reduce the number of strains without reducing the biodiversity of the whole collection. In general the nifD-nifK target proved to be more polymorphic than the rrn target. From 51 isolates of Elaeagnus frankiae, PCR-RFLP results allowed us to detect 13 identical strains, and to predict that the genomic species P8 of Akimov and Dobritsa (1992) very likely agrees with genomic species 5 of Fernandez et al. (1989). Moreover, we revealed genomic groups not yet described, as well as intraspecific variability. For Alnus frankiae, the polymorphisms shown by both the nif and the rrn PCR-RFLPs revealed three host plant species-specific subgroups inside Frankia alni. An expandable data base was created to serve as reference for future biodiversity evaluations on both culture collections and unisolated Frankia populations. It will be accessible by Internet at the International Frankia Website (http://www.unifi.it/unifi/distam/frankia/international.html).Key words: Frankia, PCR-RFLP, nifD-nifK intergenic spacer, rrn 16S-23S intergenic spacer, biodiversity, culture collections.

Polymerase chain reaction–restriction fragment length polymorphism (PCR–RFLP) for the identification of Trichinella isolates

Parasitology ◽  
1999 ◽  
Vol 118 (2) ◽  
pp. 211-218 ◽  
Author(s):  
Z. WU ◽  
I. NAGANO ◽  
E. POZIO ◽  
Y. TAKAHASHI
Keyword(s):  
Polymerase Chain Reaction ◽  
Restriction Fragment Length Polymorphism ◽  
Restriction Fragment ◽  
Length Polymorphism ◽  
Fragment Length ◽  
Fragment Length Polymorphism ◽  
Chain Reaction ◽  
Pcr Rflp ◽  
Polymerase Chain ◽  
Restriction Fragment Length

In the present study, polymerase chain reaction–restriction fragment length polymorphism (PCR–RFLP) analysis was developed to identify 5 species (Trichinella spiralis, Trichinella britovi, Trichinella nativa, Trichinella nelsoni and Trichinella pseudospiralis) and 3 phenotypes of uncertain taxonomic status (Trichinella T5, T6, and T8). Eleven restriction endonucleases were used to restrict 3 DNA fragments (1) a 2800 bp fragment of the 43 kDa excretory–secretory (E–S) protein gene, (2) a 1250 bp fragment amplified with the primer pair SB147A and (3) a 372 bp fragment amplified with the primer pair SB372A. This RFLP method allows the identification of the 8 Trichinella phenotypes as follows: T. spiralis by the HinfI or DdeI endonuclease restriction of the 2800 bp fragment; T. nativa by the RsaI restriction of the 2800 bp fragment, or by the AluI restriction of the 1250 bp fragment; T. britovi and Trichinella T8 by the AluI restriction of the 1250 bp fragments, and can be discriminated between them by the SspI restriction of the 2800 bp fragment; T. pseudospiralis by the MspI restriction of the 372 bp fragment; T. nelsoni by the HhaI or AluI restriction of the 2800 bp fragment; Trichinella T5 by the HhaI restriction of the 2800 bp fragment; Trichinella T6 by the AluI restriction of the 1250 bp fragment; and Trichinella T8 by the SspI or RsaI restriction of the 2800 bp fragment. This study reveals also an intraspecifies polymorphism in the 2800 bp and 1250 bp fragments for T. britovi, Trichinella T5 and T6.

scholarly journals Comparison of DNA Profiling between Fishes and Pork Meat using Polymerase Chain Reaction-Restriction Fragment Length Polymorphisms (PCR-RFLP) Analysis

2018 ◽  
Vol 47 (07) ◽  
pp. 1535-1540
Author(s):  
Safiyyah Shahimi ◽  
Sahilah Abd. Mutalib ◽  
Wan Sakeenah Wan Nazri ◽  
Aminah Abdullah ◽  
Norrakiah Abdullah Sani
Keyword(s):  
Polymerase Chain Reaction ◽  
Restriction Fragment ◽  
Fragment Length ◽  
Rflp Analysis ◽  
Pork Meat ◽  
Chain Reaction ◽  
Length Polymorphisms ◽  
Pcr Rflp ◽  
Polymerase Chain ◽  
Restriction Fragment Length
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