Detection of Theileria annulata by the PCR-RFLP in ticks (Acari, Ixodidae) collected from cattle in West and North-West Iran

2011 ◽  
Vol 56 (1) ◽  
Author(s):  
Mosa Tavassoli ◽  
Mohammad Tabatabaei ◽  
Bijan Nejad ◽  
Mehran Tabatabaei ◽  
Amin Najafabadi ◽  
...  
Keyword(s):  
Restriction Enzymes ◽  
Theileria Annulata ◽  
Gene Encoding ◽  
Hyalomma Anatolicum Anatolicum ◽  
North West ◽  
Blood Smears ◽  
Pcr Rflp ◽  
Single Pattern ◽  
Pcr Products ◽  
Bovine Theileriosis

AbstractThe presence of potential vectors, ticks, and susceptible hosts of bovine malignant theileriosis in all parts of Iran pose a real threat to food animal industry. The present study was conducted to determine the infection rate of ticks collected from naturally occurring bovine theileriosis in West and North-West Iran. Two hundred and thirty seven cattle suspected of suffering from theileriosis were investigated for the presence of Theileria annulata in the blood smears and any tick species on their body. In this study, 402 ticks were obtained from 99 cattle. The examination of 402 ticks by polymerase chain reaction (PCR) using primers derived from the gene encoding heat shock protein70 (Hsp70) revealed that 39.9% of Hyalomma anatolicum anatolicum, 3.5% of H. asiaticum asiaticum, and 18.2% H. anatolicum excavatum, were infected with T. annulata. The results suggest that H. a. anatolicum may play a major role in transmission of T. annulata infection in Iran. Finally, digestion of the PCR products of T. annulata with two different restriction enzymes produced only a single pattern.

scholarly journals Identification of restriction enzyme in the FSHR gene of indonesian local cattle

2021 ◽  
Vol 888 (1) ◽  
pp. 012024
Author(s):  
P W Prihandini ◽  
A Primasari ◽  
M Luthfi ◽  
D Pamungkas ◽  
A P Z N L Sari ◽  
...  
Keyword(s):  
Restriction Enzyme ◽  
Restriction Site ◽  
Follicle Stimulating Hormone ◽  
Restriction Enzymes ◽  
Future Studies ◽  
Fshr Gene ◽  
Pcr Rflp ◽  
Mapping Analysis ◽  
Pcr Products ◽  
Enzyme Mapping

Abstract The restriction enzyme is important for genotyping using the PCR-RFLP technique. Therefore, this study aims to identify the restriction enzyme mapping in the partial sequence of the follicle-stimulating hormone receptor (FSHR) gene in Indonesian local cattle. A total of 29 samples sized 306 bp, were aligned with Genbank sequence acc no. NC_032660, resulting three polymorphic sites, namely g.193G>C, g.227T>C, and g.275A>C. Furthermore, the restriction mapping analysis using the NEBcutter program V2.0 showed that no enzyme recognized the SNP g.275A>C, while the SNP g.193G>C and g.227T>C were identified by the AluI and MscI enzymes, respectively. The AluI enzyme cuts at two positions (193 bp and 243 bp) in the G allele sample producing three fragments namely 50 bp, 63 bp, and 193 bp, meanwhile, in the C allele, the AluI cuts only in position 243 bp, hence, the fragment products are 63 bp and 243 bp. In contrast, the MscI enzyme was only recognized in the T allele, producing fragments sized 77 bp and 229 bp but failed to identify the restriction site along with the PCR products in the C allele. Based on the results, the SNPs (g.193G>C and g.227T>C) and restriction enzymes (AluI and MscI) are applicable for genotyping local Indonesian cattle using the PCR-RFLP technique in future studies.

scholarly journals Detection and Differentiation of Primate α-herpesviruses by PCR

1997 ◽  
Vol 9 (3) ◽  
pp. 225-231 ◽  
Author(s):  
Daria H. Black ◽  
R. Eberle
Keyword(s):  
Pcr Primers ◽  
Restriction Pattern ◽  
Restriction Enzyme Digestion ◽  
Gene Encoding ◽  
B Virus ◽  
Pcr Rflp ◽  
Pcr Products ◽  
Unique Restriction ◽  
Polymerase Chain ◽  
Simplex Virus

A rapid method for detection and differentiation of 5 primate αpha-herpesviruses (human herpes simplex virus types 1 and 2 [HSV1, HSV2], green monkey simian agent 8, baboon herpesvirus 2 [HVP2], and macaque B virus [BV]) was developed utilizing the polymerase chain reaction (PCR). PCR primers were located in conserved regions of the gene encoding the glycoprotein B, which flanks an intervening region that is highly divergent among the 5 viruses. Amplified PCR products from the 5 viruses were readily differentiated by their unique restriction enzyme digestion patterns. No variation in digestion patterns was noted among strains of HSV1, HSV2, or HVP2. One clinical isolate of BV exhibited variation in a single restriction site, but its overall restriction pattern remained typical of BV. This method (PCR/RFLP) allowed the presence of herpesvirus DNA in clinical swabs from primates to be readily detected and the virus unambiguously identified.

scholarly journals Molecular Detection and Differentiation of Theileria lestoquardi, T. ovis and T. annulata in Blood of Goats and Ticks in Kermanshah Province, Iran

2019 ◽  
Author(s):  
Mozhgan Rahmani-Varmale ◽  
Mousa Tavassoli ◽  
Bijan Esmaeilnejad
Keyword(s):  
Nested Pcr ◽  
Molecular Detection ◽  
Ixodid Ticks ◽  
Hyalomma Anatolicum ◽  
Blood Samples ◽  
Theileria Lestoquardi ◽  
Blood Smears ◽  
Pcr Rflp ◽  
Pcr Products ◽  
Theileria Spp

Background: This study was carried out to identify Theileria spp. infections in goats and ticksin Kermanshah Prov­ince, western Iran from May–Sep 2015. Methods: For differentiation of different Theileria spp. both blood and tick samples were examined by nested PCR-RFLP. Results: Light microscopy of blood smears revealed Theileria spp. infection in 22 (5.5%), while 68 (17%) of blood samples were positive using nested PCR. Out of 68 positive samples, 85.3% (58/68) and 11.7% (8/68) were respec­tively positive for Theileria ovis and T. lestoquardi. Mixed infection was detected in 3% (2/68) cases. Overall, 420 ixodid ticks belong to seven different hard ticks species were collected from goats. Rhipicephalus turanicus 112 (26.7%), R. sanguineus 95 (22.6%), R. bursa, 91(21.7%), Hyalomma anatolicum, 55(13.1%), H. excavatum 27(6.4%), H. marginatum, 22(5.3%) and Dermacentor marginatus, 18(4.2%) were the main tick species infesting goats. The PCR products obtained from ticks were subjected to the differentiation of Theileria species. Respectively, 2 and 8 pools of H. marginatum and R. turanicus salivary glands were infected with T. ovis and T. lestoquardi. In addition, T. annulata and T. lestoquardi infection weredetected in three pools of H. anatolicum. Conclusion: This is the first report of goats and collected ticks to Theileria spp infection in Iran. The results suggest that T. ovis has a higher prevalence than T. lestoquardi. It is also postulated H. marginatum, R. turanicus and H. ana­tolicum might play an important role in the field as a vector of Theileria spp in this area.

Diversity of the desert truffle Terfezia boudieri Chatin. in southern Tunisia

2011 ◽  
Vol 57 (7) ◽  
pp. 599-605 ◽  
Author(s):  
Imed Sbissi ◽  
Faten Ghodhbane-Gtari ◽  
Mohamed Neffati ◽  
Hadda Ouzari ◽  
Abdellatif Boudabous ◽  
...  
Keyword(s):  
Its Region ◽  
Restriction Enzymes ◽  
Low Ph ◽  
Fruiting Bodies ◽  
Desert Truffle ◽  
Southern Tunisia ◽  
Pcr Rflp ◽  
Pcr Products ◽  
Novel Taxa ◽  
Terfezia Boudieri

This study reports the genetic diversity of Terfezia boudieri collected from southern Tunisia. The study was carried out using 135 truffle fruiting bodies harvested from seven different locations. Twenty-eight Terfezia claveryi fruiting bodies were also sampled from one of the seven locations. A PCR-based technique was used to amplify the internal transcribed spacer (ITS) region of the rDNA, including the ITS1–5.8S–ITS2. The PCR products were digested with the four restriction enzymes RsaI, HhaI, AluI, and HinfI. Based on the HinfI patterns, T. boudieri specimens were separated into two different haplotypes (I and II). Nucleotide sequences of some representative amplicons were also obtained. Based on the phylogenetic results, three T. boudieri genotypes could be differentiated. One sequence, SKtb1, retrieved from PCR–RFLP of haplotype I, was obtained from a low pH soil in association with Helianthemum kahiricum . Based on the results presented in the current study, this isolate may represent a novel taxa within the Terfezia genus.

Polymorphism of the exon 3 of leptin gene in Malpura sheep

2016 ◽  
Author(s):  
A.S. Meena ◽  
A. Sahoo ◽  
R. S. Bhatt ◽  
A.S. Meena
Keyword(s):  
Genetic Variants ◽  
Genomic Dna ◽  
Extraction Method ◽  
Allelic Variation ◽  
Restriction Enzymes ◽  
Allelic Frequency ◽  
Adipose Tissues ◽  
Allelic Variants ◽  
Pcr Rflp ◽  
Pcr Products

Leptin (LEP) is primarily expressed in the adipose tissues. It regulates the feed intake, energy metabolism and body composition and plays a crucial role in regulating body weight and growth in mammals. The aim of the present study was to find out an allelic variation in leptin gene of Malpura sheep. A total of 112 Malpura sheep were selected and the genomic DNA was isolated by phenol-chloroform extraction method. PCR was carried out in order to amplify the 471 bp fragment of the exon 3 coding sequence of the leptin gene. The genotyping was done by PCR-RFLP technique. PCR products were digested with three (BcnI, SsiI and OliI) restriction enzymes. For detection of allelic variants, three non-synonymous SNPs were found in Malpura sheep. The A271G and A316C loci were found mono-morphic, while T387G locus was found polymorphic. Two genetic variants (G and T) and three genotypes (GG, GT and TT) were found in Malpura sheep. The allelic frequency of G and T allele at T387G locus was found 0.82 and 0.18, respectively.

Rapid and reliable identification of rice genomes by RFLP analysis of PCR-amplified Adh genes

Genome ◽  
2001 ◽  
Vol 44 (6) ◽  
pp. 1136-1142 ◽  
Author(s):  
Song Ge ◽  
Tao Sang ◽  
Bao-rong Lu ◽  
De-yuan Hong
Keyword(s):  
Rflp Analysis ◽  
Restriction Enzymes ◽  
Pcr Primers ◽  
Specific Pcr ◽  
Adh Genes ◽  
Pcr Rflp ◽  
Restriction Sites ◽  
Pcr Products ◽  
Adh Gene ◽  
Genus Oryza

The rice genus (Oryza L.) consists of 24 species with 10 recognized genome types. With the realization of many useful genes in species of wild rice, continuous efforts have been made to understand their genomic composition and relationships. However, the identification of rice genomes has often been difficult owing to complex morphological variation and formation of allotetraploids. Here we propose a rapid and reliable method for identifying rice genomes based on the restriction sites of PCR-amplified Adh genes. The experimental procedure was as follows: (i) amplify a portion of Adh1 and Adh2 genes with the locus-specific PCR primers; (ii) digest PCR products with restriction enzymes that distinguish different genomes; and (iii) run the digested products on 1.4% agarose gel, and photograph. Using various combinations of restriction digestion of the two Adh genes, all of the rice genomes can be identified.Key words: Adh gene, genome, identification, Oryza L., PCR–RFLP.

scholarly journals Molecular surveillance and phylogenetic analysis of Theileria annulata in bovine at Baghdad city/ Iraq

2019 ◽  
Vol 43 (1) ◽  
pp. 93-101
Author(s):  
May Hameed Kawan
Keyword(s):  
Phylogenetic Analysis ◽  
Theileria Annulata ◽  
Sequencing Analysis ◽  
Local Breed ◽  
Four Seasons ◽  
Blood Smears ◽  
Significant Difference ◽  
Dna Sequencing Analysis ◽  
Bovine Theileriosis ◽  
Theileria Spp

This study is conducted to investigate Theileria spp. by traditional and molecular methods. A total of 150 blood and 50 lymph samples were collected from local breed symptomatically and asymptomatically cattle of both sexes with age ranging from less than 6 months to more than 1 year during the four seasons of 2018, in different parts at Baghdad city / Iraq. Microscopic examination of Giemsa stained blood smears revealed 39.33 %( 59/150) rate of infection with bovine theileriosis and 34 %( 17/50) positive lymph smears. Statistically no significant difference recorded between female and male: 42.04 % (37/88) and 35.48 % (22/62) respectively. Higher rate of infection 57.97 % (40/69) were recorded in more than 1 year age and 0 % in less than 6 months. 48.93 % (23/47) rate of bovine theileriosis was recorded during summer and 39.53 % (17/43) ; 37.5 % (15/40) rates were recorded during spring and autumn respectively, while the lower rate recorded in winter 20 % (4/20). DNA extraction and polymerase chain reaction (conventional PCR) were done on all cattle blood samples the result recorded that 22 out of 25 samples were positive for Theileria spp and Theileria annulata with percentage of 88 %. Also DNA sequencing analysis and genetic relationship were conducted by phylogenetic analysis.

scholarly journals Identification and differentiation among chicken’s, duck’s, quail’s, rabbit’s and turkey's meat using PCR-RFLP technique

2015 ◽  
Vol 31 (1) ◽  
pp. 101-108 ◽  
Author(s):  
S.M. Abdel-Rahman ◽  
A.M. Elmaghraby ◽  
A.S. Haggag
Keyword(s):  
Mitochondrial Dna ◽  
Cytochrome B ◽  
Fragment Length ◽  
Restriction Analysis ◽  
Restriction Enzymes ◽  
Pcr Product ◽  
Pcr Rflp ◽  
Pcr Products ◽  
Species Specific ◽  
Rflp Assay

PCR-RFLP technique was developed for identification and differentiation among chicken?s, duck?s, quail?s, rabbit?s and turkey's meat. DNA from small amount of muscles (0.05 g) was extracted and a region of mitochondrial DNA (cytochrome-b gene) in chicken, duck, quail, rabbit and turkey was amplified by PCR. Fragment length of the PCR product was 371 bp in chicken, 374 bp in duck and rabbit and 377 bp in both quail and turkey. Six nucleotides different makes it difficult to differentiate among these five species-specific meat. For differentiation, three different restriction enzymes (DdeI, MspI and TaqI) were used to digest the PCR products. Restriction analysis showed difference among chicken?s, duck?s, quail?s, rabbit?s and turkey's meat. Where, DdeI yielded two fragments (291 and 83 bp) only in rabbit?s meat. MspI yielded three fragments (221, 85 and 65 bp) in chicken?s meat and two fragments (290 and 87 bp) in both quail?s and turkey's meat. TaqI yielded three fragments (146, 134 and 94 bp) in duck?s meat and two fragments (226 and 151 bp) in quail?s meat. The use of Cytb- PCR-RFLP assay allowed a direct and fast authentication and differentiation among chicken?s, duck?s, quail?s, rabbit?s and turkey's meat.

scholarly journals Genetic Homogenity of Commerson’s Anchovy (Stolephorus commersonnii) in Segara Anakan Cilacap Central Java Inferred from PCR-RFLP Markers

2019 ◽  
Vol 7 (1) ◽  
pp. 14
Author(s):  
Agus Nuryanto ◽  
Rani Eva Dewi ◽  
Hendro Pramono
Keyword(s):  
Genetic Diversity ◽  
Restriction Enzymes ◽  
Coi Gene ◽  
Survey Method ◽  
Rflp Markers ◽  
Small Pelagic Fish ◽  
Central Java ◽  
Low Genetic Diversity ◽  
Pcr Rflp ◽  
Pcr Products

Commerson’s anchovy (Stolephorus commersonnii) is a small pelagic fish that live in a group and its existence is very abundant in Segara Anakan Cilacap. This anchovy is widely consumed by communities live around Segara Anakan. This leads to a high exploitation rate. Exploited populations generally have low genetic diversity. This study aims to evaluate genetic diversity of commerson’s anchovy population in Segara Anakan Cilacap inferred from PCR-RFLP of the cytochrome c oxidase 1 (CO1) gene. This study was conducted from January to April 2018 and used survey method by applying random sampling. As many as 30 samples of anchovy were taken. Genomic mtDNA was isolated using modified Chelex method. Partial sequences of the COI gene were amplified using a pair forward commercially available primer. The lengths of 650 base pair of the PCR products were digested with four restriction enzymes. The HindIII enzyme produces PCR-RFLP fragment with the size of 416 bp and 234 bp lengths, VspI produces 435 bp and 214 bp, CO1-TaqI produces 556 bp and 94 bp and RsaI produces 319 bp, 183 bp, and 148 bp fragments, respectively. The PCR-RFLP fragments were obtained from all samples but they produced uniform band pattern for all 30 anchovy individuals. These results indicated that the anchovy population in Segara Anakan Cilacap has monomorphic allele for all PCR-RFLP markers. Hence, it can be concluded that genetic homogenity was observed on anchovy population in Segara Anakan Cilacap as inferred from PCR-RFLP COI gene.

scholarly journals Analysis of Kinetoplast DNA from Mexican Isolates ofLeishmania (L.) mexicana

2012 ◽  
Vol 2012 ◽  
pp. 1-7 ◽  
Author(s):  
Omar Hernández-Montes ◽  
Saúl González Guzmán ◽  
Federico Martínez Gómez ◽  
Douglas C. Barker ◽  
Amalia Monroy-Ostria
Keyword(s):  
Rflp Analysis ◽  
Restriction Enzymes ◽  
The Other ◽  
Polymorphism Analysis ◽  
Specific Primers ◽  
Restriction Fragment Polymorphism ◽  
Pcr Rflp ◽  
Pcr Products ◽  
Similar Restriction ◽  
Polymerase Chain

This study analyzed DNA minicircles of Mexican isolates ofL. (Leishmania) mexicanato look for genetic differences between strains isolated from patients with diffuse cutaneous (DCL) and localized (LCL) leishmaniasis. The kDNA was analyzed using polymerase chain reaction (PCR), restriction fragment polymorphism analysis of the PCR products (PCR-RFLP) and the PCR products were sequenced. In the PCR with primers specific for the subgenusLeishmania, the Mexican isolates gave higher amplification products than the otherL. mexicanacomplex strains and with specific primers for theL. mexicanacomplex they were poorly amplified. In the PCR-RFLP analysis with theEco RV,Hae III, andMbo Iendonucleases, the Mexican isolates displayed similar restriction patterns, but different from the patterns of the other members of theL. mexicanacomplex. In the phylogenetic tree constructed, the kDNA sequences of the Mexican clones formed two groups including sequences of LCD or LCL clones, apart from the otherL. mexicanacomplex members. These results suggest that the kDNA minicircles of the Mexican isolates are more polymorphic than the kDNA of other members of theL. mexicanacomplex and have different recognition sites for the restriction enzymes used in this study.

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