Detection of Nontuberculous Mycobacterial Species from Tissue Samples of Cattle and Buffaloes by PCR and PRA (PCR-RFLP)

2021 ◽  
Author(s):  
Pallvi Slathia ◽  
Deepti Narang ◽  
Mudit Chandra
Keyword(s):  
Gene Amplification ◽  
Restriction Analysis ◽  
Polymorphism Analysis ◽  
Opportunistic Pathogens ◽  
Mycobacterial Species ◽  
Tissue Samples ◽  
Analysis Technique ◽  
Pcr Rflp ◽  
Hsp65 Gene ◽  
Polymerase Chain

Background: Nontuberculous mycobacteria are opportunistic pathogens and some of them may cause disease in humans and animals causing pulmonary infections, mastitis, lesions in respiratory tract and lymph nodes of cattle, due to which they are being recognized worldwide and also interfere with the diagnosis of bovine tuberculosis. Methods: The present study was conducted for detection of nontuberculous mycobacterial species (NTM) in tissue samples (with and without tubercle lesions) in cattle and buffaloes from postmortem hall GADVASU, Ludhiana. Polymerase Chain Reaction and PCR-RFLP which involved hsp65 gene amplification (439 bp) and restriction analysis of amplified product was performed on 30 tissue samples for detection of nontuberculous mycobacterial species. Result: Three out of 30 samples showed hsp65 gene amplification and 2 were identified as M. kansasii using restriction analysis technique and one could not be identified as the RFLP patterns was different from other known PCR-RFLP profiles. NTM such as M. kansasi may cause infection in animals and PRA (PCR-Restriction Fragment Length Polymorphism Analysis) technique was found to be a rapid tool for identification and differentiation of NTM upto species level.

Rapid Molecular Typing of Methicillin-ResistantStaphylococcus aureusby PCR-RFLP

2001 ◽  
Vol 22 (5) ◽  
pp. 294-298 ◽  
Author(s):  
Thomas A. Wichelhaus ◽  
Klaus-Peter Hunfeld ◽  
Boris Böddinghaus ◽  
Peter Kraiczy ◽  
Volker Schàfer ◽  
...  
Keyword(s):  
Fragment Length Polymorphism ◽  
Restriction Analysis ◽  
Protein A ◽  
Hypervariable Region ◽  
Epidemiological Surveillance ◽  
Typing Method ◽  
Rapid Pcr ◽  
Pcr Rflp ◽  
Polymerase Chain ◽  
Rflp Typing

AbstractObjective:To establish a new, rapid, and reliable genotypic fingerprinting technique for methicillin-resistantStaphylococcus aureus(MRSA) typing in routine epidemiological surveillance.Design:The method is based on polymerase chain reaction (PCR) restriction fragment-length polymorphism (RFLP) followingHaeII digestion of simultaneously amplified parts of the protein A gene, the coagulase gene, and the hypervariable region adjacent tomecA. A total of 46 MRSA initial isolates were analyzed, including 14 isolates from five countries; the six German epidemic strains; 16 isolates from the Frankfurt metropolitan area, which were known to be heterogeneous by pulsed-field gel electrophoresis (PFGE); and 10 isolates obtained during three epidemics, all of which displayed an identical genotype.Results:Restriction analysis by PCR-RFLP permitted discrimination of 10 of 14 international isolates, all six German epidemic strains, and 15 of 16 national isolates. It also confirmed the homogeneous character of the 10 outbreak isolates.Conclusions:This new and rapid PCR-RFLP typing method is an attractive tool in routine epidemiological surveillance. Its impressive characteristics are ease of performance and interpretation, while at the same time guaranteeing good discriminatory power, reproducibility, and typeability.

scholarly journals Genetic polymorphism analysis of 5' untranslated region of thyroglobulin gene in Bali cattle (Bos javanicus) from three different regions of Indonesia

2017 ◽  
Vol 42 (3) ◽  
pp. 175
Author(s):  
S. Anwar ◽  
A.C. Putra ◽  
A.S. Wulandari ◽  
P. P. Agung ◽  
W.P.B. Putra ◽  
...  
Keyword(s):  
Genetic Polymorphism ◽  
Untranslated Region ◽  
Candidate Snps ◽  
Polymorphism Analysis ◽  
Bos Javanicus ◽  
Pcr Rflp ◽  
Thyroglobulin Gene ◽  
Polymerase Chain ◽  
Reference Sequences ◽  
Bali Cattle

The g.422C>T nucleotide variations in the 5’ untranslated region (5’UTR) of TG gene (called as TG5) has been reported to be associated with level in intramuscular fat (IMF) content or marbling in beef cattle. The objective of this study was to confirm genetic polymorphism of TG5 gene in Bali cattle populations from three different regions as the main resources of Bali cattle in Indonesia. A total of 200 head of Bali cattle have been performed genotyping on TG5 gene using polymerase chain reaction-restriction fragment lenght polymorphism (PCR-RFLP) method and sequence analysis. Results of the study confirmed that TG5 was monomorphic in Bali cattle wherever their origin regions. Moreover, nine candidate SNPs were detected within 5’UTR of TG gene in Bali cattle compared to Genbank reference sequences, although no SNP variations among Bali cattle sample studied. The new other genetic markers within an entire TG gene suggested to be explored and verified for their polymorphisms in Bali cattle. The nine candidate SNPs were also required further verification and validation in a larger sample to be regarded as new SNPs between Bali cattle and Genbank reference sequences.

scholarly journals A quick PCR-based method for identification of Melolontha melolontha and Melolontha hippocastani (Coleoptera: Scarabaeidae

2018 ◽  
Vol 29 (3) ◽  
pp. 141-145
Author(s):  
Anna Tereba ◽  
Marzena Niemczyk
Keyword(s):  
Insect Pests ◽  
Economic Losses ◽  
Morphological Identification ◽  
Effective Management ◽  
Severe Damage ◽  
Tissue Samples ◽  
Melolontha Melolontha ◽  
Pcr Rflp ◽  
The Common ◽  
Polymerase Chain

The common cockchafer (Melolontha melolontha) and the forest cockchafer (Melolontha hippocastani) are among the most destructive insect pests in many European countries. Larvae feed on the roots of numerous plant species, thus inflicting severe damage and heavy economic losses. The two species are often discussed together because they are difficult to distinguish during the larval stage.However, they differ slightly in ecology and development. The aim of this study was to develop a quick PCR-RFLP (polymerase chain reaction-restriction fragment length polymorphism) method for easily identifying the two Melolonthaspecies through tissue samples or larvae, when reliable morphological identification is lacking. The strength of the method was tested on 43 M. melolonthaand 37 M. hippocastani individuals. We demonstrate that the technique is rapid and inexpensive, with strong implications for the effective management of these insect pests.

scholarly journals The Frequencies of Allele Distribution of CYP2C9 and CYP2C19 Gene Polymorphisms in Healthy Papuan Population, Indonesia

2021 ◽  
Vol 20 (3) ◽  
Author(s):  
Syahrul Tuba ◽  
Zullies Ikawati ◽  
Mustofa
Keyword(s):  
Ethnic Group ◽  
Gene Polymorphisms ◽  
Fragment Length Polymorphism ◽  
Polymorphism Analysis ◽  
Allele Distribution ◽  
Pcr Rflp ◽  
Cyp2c19 Gene ◽  
Hardy Weinberg Equilibrium ◽  
Polymorphism Rate ◽  
Polymerase Chain

This study's objective was to determine the distribution of allele frequencies of CYP2C9 and CYP2C19 gene polymorphisms among the Papuan population, known as the second-largest ethnic group in Indonesia. According to recent research, there is a decrease in CYP2C9 and CYP2C19 produced by humans globally, including in Indonesia. These gene polymorphisms aid in the transmission of various endogenous and exogenous drugs in the human body. Material and Methods: A sum of 99 subjects, comprising 73 male and 26 female subjects aged 20-30 years, were used for this research. PCR-RFLP (polymerase chain reaction-restriction fragment length polymorphism) analysis using AvaII, NsiI, and SfaNI enzymes tested for the genotypes CYP2C9 and CYP2C19 administered. The distribution of genotypes was calculated in the population (P<0.05) using the Hardy-Weinberg equilibrium. The Faculty of Medicine Gadjah Mada University's Medical and Health Research Ethics Committee (MHREC) accepted this research with written consent. The results revealed that in Papua subjects, CYP2C9*2 (rs1799853) and CYP2C19*17 (rs12248560) alleles were absent while in 17 percent of the population CYP2C9*3 (rs1057910) allele frequency was. In conclusion, CYP2C9*3 has the highest polymorphism rate in Indonesia, with the absence of CYP2C9*2 and CYP2C19*17. Therefore, genetic drift can occur within this ethnic group. Keywords: Genotyping; Papuan ethnic; Pharmacogenetics; Polymorphisms

scholarly journals Cytauxzoon felis and 'Candidatus Mycoplasma haemominutum' coinfection in a Brazilian domestic cat (Felis catus)

2013 ◽  
Vol 22 (2) ◽  
pp. 289-291 ◽  
Author(s):  
Leticia Mendes Pupio Maia ◽  
Aloysio de Mello Figueiredo Cerqueira ◽  
Daniel de Barros Macieira ◽  
Aline Moreira de Souza ◽  
Namir Santos Moreira ◽  
...  
Keyword(s):  
Ribosomal Rna ◽  
Bacterial Species ◽  
Molecular Techniques ◽  
Domestic Cat ◽  
Polymorphism Analysis ◽  
Sequencing Analysis ◽  
Gene Encoding ◽  
Cytauxzoon Felis ◽  
Pcr Rflp ◽  
Polymerase Chain

This article describes the first detection of Cytauxzoon felis, using molecular techniques, in a naturally infected domestic cat from Brazil, South America. Coinfection with 'CandidatusMycoplasma haemominutum' was also found. The molecular identification of the piroplasmid species was performed by Polymerase Chain Reaction (PCR) and sequencing analysis. A 284 pb fragment of the gene encoding the 18S ribosomal RNA region was amplified and showed 99% identity with other C. felis strains from North America. In addition, PCR-RFLP (restriction fragment length polymorphism) analysis, which amplifies a 595 bp fragment of the gene encoding 16S ribosomal RNA of some bacterial species, identified the co-infecting species as 'Candidatus M. haemominutum'.

scholarly journals Restriction fragment length polymorphism analysis of isolates of infectious bursal disease viruses from Turkey

2012 ◽  
Vol 48 (No. 12) ◽  
pp. 359-362 ◽  
Author(s):  
G. Ozbes ◽  
Ertas HB ◽  
A. Muzo
Keyword(s):  
Rflp Analysis ◽  
Infectious Bursal Disease ◽  
Polymorphism Analysis ◽  
Rt Pcr ◽  
Vp2 Gene ◽  
Infectious Bursal Disease Viruses ◽  
Pcr Rflp ◽  
Bursal Disease ◽  
Polymerase Chain ◽  
Rflp Assay

Infectious bursal disease Virus (IBDV) specific reverse transcriptase/polymerase chain recation (RT/PCR) positive 40 broiler bursa fabricius samples obtained from a commercially reared flock were investigated for genetic diversity by PCR-RFLP assay. The assay amplifies a 743 bp fragment of the IBDV VP2 gene. The RFLP profiles of 40 of these positive samples were determined using the enzyme MboI. Most of the viruses had the same RFLP with the MboI enzyme. RFLP analysis of the isolates produced two different band profiles. The results of this study showed that little genetic heterogeneity exists among IBDV strains in a infected flock.

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