The gp63 gene locus, a target for genetic characterization of Leishmania belonging to subgenus Viannia

Parasitology ◽  
1998 ◽  
Vol 117 (1) ◽  
pp. 1-13 ◽  
Author(s):  
K. VICTOIR ◽  
A. L. BAÑULS ◽  
J. AREVALO ◽  
A. LLANOS-CUENTAS ◽  
R. HAMERS ◽  
...  
Keyword(s):  
Gene Locus ◽  
Genetic Characterization ◽  
Restriction Analysis ◽  
Pcr Amplification ◽  
Rflp Analysis ◽  
Restriction Enzymes ◽  
Pcr Rflp ◽  
Natural Isolates ◽  
Phenotype Diversity

In the present study the gp63 gene locus was used as a target for genetic characterization of Leishmania parasites by 2 methods: (i) RFLP analysis with several restriction enzymes (gp63–RFLP), and (ii) intra-genic PCR amplification coupled with restriction analysis (PCR–RFLP). Both methods were applied to a large number of natural isolates belonging to 4 species of the subgenus Viannia, namely L. (V.) braziliensis, L. (V.) peruviana, L. (V.) guyanensis and L. (V.) lainsoni: reference stocks of subgenus Leishmania were included as outgroups. Multilocus isoenzyme typing (MLEE) was used as a reference. On the one hand gp63–RFLP evidenced an extensive polymorphism and revealed specific markers for subgenus, species and geographical populations: congruence with MLEE was demonstrated statistically. The particular interest of gp63–RFLP was illustrated by infra-specific polymorphism, because of the possible relationship with phenotype diversity. On the other hand intra-genic amplification was less resolutive than gp63–RFLP, but also allowed discrimination of the 2 subgenera (PCR alone) and all the species tested in the subgenus Viannia (PCR–RFLP). PCR–RFLP presents an important operational advantage as it allows genetic characterization of minute amounts of parasites, using Leishmania specific primers. The polymorphism revealed by gp63–RFLP and PCR–RFLP illustrates the very high genomic and genetic plasticity of gp63 genes.

scholarly journals Detection and characterization of Leptospira spp. isolated from aborted bovine clinical samples

2011 ◽  
Vol 81 (1) ◽  
pp. 21-25 ◽  
Author(s):  
Hassan Momtaz ◽  
Saadat Moshkelani
Keyword(s):  
Public Health Problem ◽  
Rflp Analysis ◽  
Restriction Enzymes ◽  
Clinical Samples ◽  
Dairy Herds ◽  
Important Public Health Problem ◽  
Pcr Rflp ◽  
Beef Herds ◽  
Bovine Abortion

Leptospira is recognized as an important public health problem worldwide, especially in tropical countries, and is a common cause of abortion in dairy and beef herds. The aim of the present study was to detect and characterize Leptospira as the causative agent of abortion in cattle using a PCR-RFLP in Chaharmahal va Bakhtiari and Isfahan provinces, Iran. A total of 220 bovine aborted foetuses and 120 vaginal discharges from an aborted calf were collected from 64 commercial dairy herds. After isolation of 60 Leptospira spp. from samples, RFLP analysis was carried out with HindIII and HaeIII restriction enzymes in reference strains and isolated for characterization. In a total of 340 specimens, 46 (20.9%) and 14 (11.66%) were identified positive for Leptospira spp. from aborted bovine foetuses and vaginal discharges, respectively. The present results also suggest that L. interrogans serovar hardjo has the highest prevalence in the region under study and L. hardjo is a major pathogen causing bovine abortion in Chaharmahal va Bakhtiari and Isfahan provinces of Iran.

Molecular genetic characterization of the cervid strain (‘northern form’) ofEchinococcus granulosus

Parasitology ◽  
1994 ◽  
Vol 109 (2) ◽  
pp. 215-221 ◽  
Author(s):  
J. Bowles ◽  
D. Blair ◽  
D. P. McManus
Keyword(s):  
Genetic Characterization ◽  
Molecular Genetic ◽  
Rflp Analysis ◽  
Domestic Sheep ◽  
Rflp Pattern ◽  
Its1 Region ◽  
Pcr Rflp ◽  
Mitochondrial Nd1 ◽  
Molecular Genetic Characterization

SUMMARYWe have used a number of molecular genetic approaches to characterize the cervid strain (‘northern form’) ofEchinococcus granulosus.PCR–RFLP analysis of the nuclear ITS1 region of the rDNA repeat can readily distinguish the cervid form from other strains ofE. granulosus. The complexity of the RFLP patterns obtained suggests that a number of distinct ITS1 types are present in this strain which may represent an inter-strainE. granulosushybrid. Mitochondrial CO1 sequence of the cervid genotype was ambiguous at 18 positions and closely resembles a cluster of previously characterizedE. granulosusgenotypes, G1 (common, domestic sheep)/G2 (Tasmanian sheep)/G3 (buffalo). In contrast, mitochondrial ND1 sequence, although unique, suggests that the cervid form is most similar to strains represented by the G6 (camel)/G7 (pig) genotypes. We assume that the CO1 and ND1 sequences obtained for the cervid genotype are linked in a single mitochondrial genome although this is difficult to explain if conventional molecular genetics of mitochondrial DNA are assumed. Based on its unique ND1 sequence and ITS1 PCR–RFLP pattern, the cervid strain appears to represent a distinct genotype (designated G8) ofE. granulosus.

scholarly journals Characterization of Ascaris from Ecuador and Zanzibar

2014 ◽  
Vol 89 (4) ◽  
pp. 512-515 ◽  
Author(s):  
A.M. Sparks ◽  
M. Betson ◽  
G. Oviedo ◽  
C. Sandoval ◽  
P.J. Cooper ◽  
...  
Keyword(s):  
Fragment Length Polymorphism ◽  
Rflp Analysis ◽  
Its Region ◽  
Restriction Enzymes ◽  
Zoonotic Transmission ◽  
Pcr Rflp ◽  
Double Digestion ◽  
Polymerase Chain ◽  
Shed Light

AbstractTo shed light on the epidemiology of ascariasis in Ecuador and Zanzibar, 177 adult worms retrieved by chemo-expulsion from either people or pigs were collected, measured and subjected to polymerase chain reaction–restriction fragment length polymorphism (PCR-RFLP) analysis of the ribosomal internal transcribed spacer (ITS) region. Upon double digestion with RsaI and HaeIII, PCR-RFLP analysis revealed the presence of A. lumbricoides in people and A. suum in pigs in Ecuador. In contrast, while there are no pigs on Zanzibar, of the 56 worms obtained from people, one was genotyped as A. suum. No additional genetic variation was detected upon further PCR-RFLP analysis with several other restriction enzymes. Upon measurement, worm mass and length differed by location and by species, A. suum being lighter and longer. While there is no evidence to suggest zoonotic transmission in Ecuador, an enduring historical signature of previous zoonotic transmission remains on Zanzibar.

Characterization of Fusarium spp. isolates by PCR-RFLP analysis of the intergenic spacer region of the rRNA gene (rDNA)

2006 ◽  
Vol 106 (3) ◽  
pp. 297-306 ◽  
Author(s):  
A. Llorens ◽  
M.J. Hinojo ◽  
R. Mateo ◽  
M.T. González-Jaén ◽  
F.M. Valle-Algarra ◽  
...  
Keyword(s):  
Intergenic Spacer ◽  
Rflp Analysis ◽  
Rrna Gene ◽  
Fusarium Spp ◽  
Intergenic Spacer Region ◽  
Pcr Rflp

scholarly journals Molecular Characterization of Pathogenicity Locus (PaLoc) and tcdC Genetic Diversity Among tcdA+B+ Clostridioides Difficile Clinical Isolates in Tehran, Iran

2020 ◽  
Author(s):  
Mansoor Kodori ◽  
Zohreh Ghalavand ◽  
Abbas Yadegar ◽  
Gita Eslami ◽  
Masoumeh Azimirad ◽  
...  
Keyword(s):  
Genetic Diversity ◽  
Molecular Characterization ◽  
Disease Severity ◽  
Genetic Variants ◽  
Rflp Analysis ◽  
Clostridioides Difficile ◽  
Pcr Rflp ◽  
The Common ◽  
Healthcare Associated

Abstract Background: Clostridioides difficile is the main cause of healthcare-associated diarrhea worldwide. It is proposed that certain C. difficile toxinotypes with distinct pathogenicity locus (PaLoc) variants are associated with disease severity and outcomes. Additionally, few studies have described the common C. difficile toxinotypes, and also little is known about the tcdC variants in Iranian isolates. We characterized the toxinotypes and the tcdC genotypes from a collection of Iranian clinical C. difficile tcdA+B+ isolates with known ribotypes (RTs).Methods: Fifty C. difficile isolates with known RTs and carrying the tcdA and tcdB toxin genes were analyzed. Toxinotyping was carried out based on a PCR-RFLP analysis of a 19.6 kb region encompassing the PaLoc. Genetic diversity of the tcdC gene was determined by the sequencing of the gene.Results: Of the 50 C. difficile isolates investigated, five distinct toxinotypes were recognized. Toxinotypes 0 (33/50, 66%) and V (11/50, 22%) were the most frequently found. C. difficile isolates of the toxinotype 0 mostly belonged to RT 001 (12/33, 36.4%), whereas toxinotype V consisted of RT 126 (9/11, 81.8%). The tcdC sequencing showed six variants (35/50, 70%); tcdC-sc3 (24%), tcdC-A (22%), tcdC-sc9 (18%), tcdC-B (2%), tcdC-sc14 (2%), and tcdC-sc15 (2%). The remaining isolates were wild-types (15/50, 30%) in the tcdC gene.Conclusions: The present study demonstrates that the majority of clinical tcdA+B+ isolates of C. difficile frequently harbor tcdC genetic variants. We also found that the RT 001/ toxinotype 0 and the RT 126/ toxinotype V are the most common types among Iranian isolates. Further studies are needed to investigate the putative association of various tcdC genotypes with CDI severity and its recurrence.

scholarly journals Genetic Characterization of Thyroglobulin and Leptin Genes in Pasundan Cattle at West Java

2019 ◽  
Vol 43 (1) ◽  
Author(s):  
Widya Pintaka Bayu Putra ◽  
Saiful Anwar ◽  
Syahruddin Said ◽  
Romanos Albert Adhitya Indarto ◽  
Putri Wulandari
Keyword(s):  
Genetic Characterization ◽  
Rare Allele ◽  
Basic Information ◽  
Dominant Allele ◽  
West Java ◽  
Pcr Rflp ◽  
Molecular Selection ◽  
Lep Gene ◽  
Informative Content

The Thyroglobulin (TG) and Leptin (LEP) genes are two candidate genes that widely used for molecular selection to improve carcass traits in beef cattle. This research was carried out to identify the genetic characterization of TG and LEP genes from 47 heads of Pasundan cows at West Java using PCR-RFLP method. Research shows that TG gene of Pasundan cattle is monomorphic with C allele as the dominant allele (1.00). However, LEP gene of Pasundan cattle is polymorphic with C allele as the dominant allele (0.98) and T as the rare allele (0.02). The polymorphic informative content (PIC) and numberof effective allele (ne) values in the LEP gene in the animal studied were 0.04 and 1.04 respectively. It was concluded that TG/BstYI and LEP/Sau3AI gene in the present study can not be used as molecular selection in Pasundan cattle. These results are important as the basic information for preparing the molecular selection program in the future.

scholarly journals Identification of Pneumococcal Serotypes by PCR–Restriction Fragment Length Polymorphism

Diagnostics ◽  
2019 ◽  
Vol 9 (4) ◽  
pp. 196 ◽  
Author(s):  
García-Suárez ◽  
González-Rodríguez ◽  
Cima-Cabal ◽  
Yuste ◽  
Vazquez ◽  
...  
Keyword(s):  
Restriction Fragment Length Polymorphism ◽  
Restriction Fragment ◽  
Length Polymorphism ◽  
Dna Sequences ◽  
Fragment Length ◽  
Fragment Length Polymorphism ◽  
Rflp Analysis ◽  
Restriction Enzymes ◽  
Pcr Rflp ◽  
Restriction Fragment Length

Streptococcus pneumoniae shows more than 90 capsular serotypes that can be distinguished by their reactivity against antisera. The main objective of this work was the development of a molecular method for serotyping without the use of antisera. A computer program containing an algorithm was used to search in a database for potentially useful enzymes for Restriction Fragment Length Polymorphism-RFLP typing, in order to maximize the discrimination between different serotypes. DNA sequences of 90 serotypes for the region between dexB and aliA genes were compiled, and a computer screening of restriction enzymes was performed. The wzg–wzh–wzd–wze region and Sse9I restriction predicted unique PCR-RFLP patterns for 39 serotypes and eight serogroups. A second restriction enzyme resolved fragment specific patterns for 25 serotypes. The method was tested with 98 serotype-unknown clinical isolates. PCR-RFLP analysis deduced correct serotypes that were confirmed by Quellung reaction for 78.5% of the isolates.

PCR-based genetic identification of marlin and other billfish

1998 ◽  
Vol 49 (5) ◽  
pp. 383 ◽  
Author(s):  
B. H. Innes ◽  
P. M. Grewe ◽  
R. D. Ward
Keyword(s):  
Mitochondrial Dna ◽  
Genetic Test ◽  
Rflp Analysis ◽  
Restriction Enzymes ◽  
Genetic Identification ◽  
Dna Molecule ◽  
Pcr Rflp ◽  
D Loop ◽  
Specific Variation ◽  
Species Specific

A genetic test was developed for the identification of the six species of billfish found in Australian waters (black marlin, Indo–Pacific blue marlin, striped marlin, Indo–Pacific sailfish, shortbill spearfish and broadbill swordfish). The test was based on the PCR–RFLP analysis of a 1400 bp region of the mitochondrial DNA molecule, the d-loop, using four restriction enzymes (Hinf I, Rsa I and Sau3A I andTaq I). A total of 33 composite haplotypes were observed among 160 fish; all were species-specific. Three of the species—black marlin, striped marlin and broadbill swordfish—showed sufficient intra-specific variation to be useful in population structure analyses.

Studies on TLR2 gene variants and their association with milk yield and milk quality traits in Bos indicus (Deoni) cattle

2017 ◽  
Author(s):  
U. T. Mundhe ◽  
D. N. Das ◽  
R. S. Gandhi ◽  
P. Divya
Keyword(s):  
Milk Yield ◽  
Bos Indicus ◽  
Restriction Enzymes ◽  
Milk Quality ◽  
Quality Traits ◽  
Exon 2 ◽  
Allelic Frequencies ◽  
Tlr2 Gene ◽  
Pcr Rflp

Present study molecular characterization of exon 2 of TLR2 gene and its association with milk yield and milk quality traits in 104 Deoni cattle using PCR- RFLP technique was done. Polymorphism was observed through HaeIII, HhaI and EcoRV restriction enzymes in Created Restriction Site (CRS) exon 2-1, CRS exon 2-5 and exon 2-1 by PCR- RFLP, respectively. In CRS exon 2-1 allelic frequencies were observed as 0.793 for A and 0.206 for B alleles and that of genotypic frequencies were 0.58 and 0.41 for genotypes AA and AB. In CRS exon 2-5, two genotypes viz., AC and CC with corresponding allelic frequencies were observed as 0.221 for A and 0.778 for C allele and that of genotypic frequencies observed were 0.44 and 0.55 for AC and CC genotypes respectively. TLR2 exon 2-1 exhibited two alleles G and T with frequencies of 0.134 and 0.865 and their Corresponding genotypic frequencies were 0.009, 0.25 and 0.74for GG, GT and TT genotypes respectively. Higher count of somatic cells (SCC) in TT homozygous and TG heterozygous genotypes, and lower in GG homozygous genotypes were observed in exon 2-1. Strongly significant (P£0.01) effect for least squares means of Test Day milk yield (TDMY) and Somatic Cell Count of CRS exon 2-1 were observed.

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