scholarly journals Molecular Typing of Methicillin ResistantStaphylococcus aureusClinical Isolates on the Basis of Protein A and Coagulase Gene Polymorphisms

2014 ◽  
Vol 2014 ◽  
pp. 1-11 ◽  
Author(s):  
Nancy Younis Omar ◽  
Hala Abdel Salam Ali ◽  
Reem Abdel Hameed Harfoush ◽  
Engy Hamdy El Khayat
Keyword(s):  
Molecular Typing ◽  
Gene Polymorphisms ◽  
Protein A ◽  
Rflp Analysis ◽  
Restriction Digestion ◽  
Banding Patterns ◽  
Pcr Rflp ◽  
Pcr Products ◽  
Mrsa Strains

Increased frequency of methicillin-resistantStaphylococcus aureus(MRSA) in hospitalized patients requires rapid and reliable characterization of isolates for control of MRSA spread in hospitals. This study evaluated polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) as a molecular typing technique for MRSA strains on the basis of protein A (spa) and coagulase (coa) gene polymorphisms to verify their ability in assessing the relatedness of isolates. Seventy-five MRSA isolates, from different ICUs of Alexandria University Main Hospital, were characterized using antibiotyping and PCR-RFLP analysis ofcoaandspagenes. Thirty-two antibiotypes were identified.coagene PCR generated 3 types and 10 subtypes of band patterns.HaeIIIrestriction digestion of amplifiedcoagene products produced 5 major banding patterns and 12 subtypes.spagene PCR products generated 4 major and 11 minor types, and theirHaeIIrestriction digestion showed 5 major and 12 minor banding patterns. The combinedcoaandspaRFLP patterns generated 22 combined R types. Typing usingcoaPCR and PCR-RFLP had the same discriminatory index (DI) value (0.64), which was comparable to that of bothspaPCR and PCR-RFLP techniques (0.68). The combined grouping increased the DI value to 0.836. The current study revealed that testing for multiple gene polymorphisms is more useful for local epidemiologic purposes.

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 Phylogenetic Relationships of Local Durian Species based on Morphological Characteristics and PCR-RFLP Analysis of the Ribosomal Internal Transcribed Spacer (ITS) DNA

2016 ◽  
Vol 8 (3) ◽  
pp. 362
Author(s):  
Fidia Fibriana ◽  
Lutfia Nur Hadiyanti
Keyword(s):  
Phylogenetic Relationships ◽  
Internal Transcribed Spacer ◽  
Morphological Characteristics ◽  
Biology Education ◽  
Rflp Analysis ◽  
Restriction Pattern ◽  
Internal Transcribed Spacers ◽  
Central Java ◽  
Pcr Rflp ◽  
Pcr Products

<p>In this study, twenty local durian accessions obtained from Central Java in situ collection were characterized using the morphological characteristics and the restriction patterns which generated from the region spanning the internal transcribed spacers ITS LEU and ITS 4. Morphological characteristics of durian leaf, stem, tree, and fruit showed variations for the different accessions, whereas polymerase chain reaction (PCR) products of ribosomal DNA region showed a low length of variation. The size of the PCR products and the restriction analyses with the restriction endonucleases Bsp1431yielded a restriction pattern for each accessions. The results of this study can be utilized by local durian farmers as a preliminary reference for durian propagation. The data obtained need to be supported by further research using the other molecular markers to obtain more accurate data. The clear identity of durian species can help the management of propagation systems by farmers to get superior local durian.</p><p><strong>How to Cite</strong></p><p>Fibriana, F., &amp; Hadiyanti, L. N. (2016). Phylogenetic Relationships of Local Durian Species based on Morphological Characteristics and PCR-RFLP Analysis of the Ribosomal Internal Transcribed Spacer (ITS) DNA. <em>Biosaintifika: Journal of Biology &amp; Biology Education</em>, 8(3), 362-370. </p>

scholarly journals Sex Determination During Inflorescence Bud Differentiation in Monoecious Pistacia chinensis Bunge

Forests ◽  
2019 ◽  
Vol 10 (3) ◽  
pp. 202 ◽  
Author(s):  
Qian Bai ◽  
Chenyi Zhu ◽  
Xia Lei ◽  
Tao Cao ◽  
Shuchai Su ◽  
...  
Keyword(s):  
Sex Determination ◽  
Early Stage ◽  
Sex Differentiation ◽  
Banding Patterns ◽  
Vegetative Period ◽  
Pcr Products ◽  
Polymerase Chain ◽  
Sex Determination Mechanisms

Pistacia chinensis Bunge is widely acknowledged to be dioecious, but rare monoecious individuals have been found. However, the origin of monoecism and the sex differentiation of different sex types remain intriguing questions. Here, sex expressions were explored by identification of sex-associated DNA markers, determination of the sex stability after grafting, and histological characterization of inflorescence bud development using anatomical analysis. The results showed that (1) although polymorphisms among individuals existed, the banding patterns of Polymerase Chain Reaction (PCR) products for different sex types on the same monoecious tree were consistent; (2) the sex expressions of grafted trees were not consistent with those of scions, indicating that monoecism probably did not originate from a stable bud mutation; and (3) both males and females underwent a bisexual period, then the stamen primordia in female buds degenerated into the second round tepals, while the pistil primordia in male buds gradually disappeared. During the sex differentiation phase, female buds were spindle-shaped, while the male buds were full teardrop-shaped, and male buds were bigger than female buds. Taken together, no sex-associated DNA marker was found, sex expressions were unstable after grafting, and the alternative sex organs appeared in the early stage of sex differentiation, suggesting that sex determination occurred during floral development instead of the early vegetative period. These results indicated that the sex expressions may be affected by environmental factors, increasing the understanding of sex determination mechanisms in P. chinensis and other species.

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.

scholarly journals Estrus Behaviour and Reproductive Traits of Pulawska Gilts Associated with Selected Gene Polymorphisms

2021 ◽  
Vol 21 (1) ◽  
pp. 159-172
Author(s):  
Marek Babicz ◽  
Marcin Pastwa ◽  
Magdalena Szyndler-Nędza ◽  
Anna Kozubska-Sobocińska ◽  
Barbara Danielak-Czech ◽  
...  
Keyword(s):  
Genetic Structure ◽  
Gene Polymorphisms ◽  
Rflp Analysis ◽  
Reproductive Traits ◽  
The Other ◽  
Molecular Analyses ◽  
Native Breed ◽  
Hair Roots ◽  
Pcr Rflp ◽  
Pcr Conditions

Abstract Searching for the associations between the gene polymorphism and the reproductive traits is essential in defining the genetic native breed specificity, which distinguishes them from the other breeds. The aim of our study was to determine the associations between mutations in the PRL, PRLR, PTGS2, FUT1 genes and sexual and periparturient activity in native Pulawska gilts. The analysis included 72 animals which gave birth to the first litter. Evaluation of the productive value of gilts accounted for indicators of sexual and periparturient activity as well as reproductive traits. The biological material for molecular analyses was obtained from the hair roots of the gilts. The genotype was verified by PCR RFLP analysis. The primers and PCR conditions were determined on the basis of available literature data. Statistically significant differences (P≤0.05) were found at the PRL locus: gilts of AA genotypes (Ins/Ins) at the PRL locus were characterised by longest farrowing duration compared to gilts of AB genotype (P≤0.05). The analysis of PRLR gene showed that gilts of TT genotype revealed a tendency for later occurrence of estrus signs (first and second estrus) and for the markedly longest farrowings (P≤0.05). With regard to PTGS2 and FUT1 loci, no significant differences were found in the parameters of sexual and periparturient activity of the gilts. However, gilts of FUT1 GG genotype gave birth to and reared the largest first litters (P≤0.05). The results of the studies expand the knowledge about the genetic structure and productivity of Pulawska gilts.

scholarly journals Molecular Identification and VOMs Characterization of Saccharomyces cerevisiae Strains Isolated from Madeira Region Winery Environments

Processes ◽  
2020 ◽  
Vol 8 (9) ◽  
pp. 1058 ◽  
Author(s):  
Mariangie Castillo ◽  
Emanuel da Silva ◽  
José S. Câmara ◽  
Mahnaz Khadem
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Solid Phase ◽  
Rflp Analysis ◽  
Selective Adaptation ◽  
Internal Transcribed Spacers ◽  
Gas Chromatography Mass Spectrometry ◽  
Pcr Rflp ◽  
Volatile Organic ◽  
Major Chemical

The quality and typical characteristic of wines depends, among other factors, on the volatile organic metabolites (VOMs) that are biosynthesized by yeasts, mainly Saccharomyces cerevisiae species. The yeast strain influences the diversity and proportions of the VOMs produced during the fermentation process, as the genetic predisposition of the strains is a by-product of selective adaptation to the ecosystem. The present work reports the characterization of S. cerevisiae strains isolated from grape must, used in the Demarcated Region of Madeira (DRM) for winemaking. Yeast species were identified by amplification and by restriction fragment length polymorphism (RFLP) analysis of the region 5.8S-internal transcribed spacers (PCR-RFLP of 5.8S-ITS) of ribosomal DNA (rDNA). The strains identification was performed by analyzing the RFLP pattern of mitochondrial DNA (RFLP-mtDNA). The representative strains were selected for the characterization of the volatile profile through headspace solid-phase microextraction (HS-SPME) followed by gas chromatography-mass spectrometry (GC-MS) analysis. A total of 77 VOMs were identified. Higher alcohols, esters, and fatty acids were the major chemical families representing 63%, 16%, and 9%, respectively, in strain A and 54%, 23%, and 15% in strain B. The results indicate the influence of the strain metabolism in the production of VOMs, many of which probably participate in the aroma of the corresponding wines.

Association of polymorphism at intron 2 of FABP3 Gene with milk production traits in Sahiwal and Karan Fries cattle

2018 ◽  
Author(s):  
Alok Kumar Yadav ◽  
Anupama Mukherjee ◽  
Suchit Kumar
Keyword(s):  
Milk Production ◽  
Rflp Analysis ◽  
Production Traits ◽  
Milk Production Traits ◽  
Pcr Rflp ◽  
Sahiwal Cattle ◽  
Pcr Products ◽  
The Mean ◽  
Intron 2 ◽  
Karan Fries

PCR-RFLP analysis of PCR products were carried out using Aci I / SSi I for 100 Sahiwal and 115 Karan Fries cattle. In Sahiwal cattle and Karan Fries cattle, 438bp has three genotypes AA (438), AB (438+299+139 bp) and BB (299+139 bp). In Sahiwal cattle these genotypes are highly significant for FL305DPY but in Karan Fries cattle these genotypes are highly significant for FL305DMY, FLTMY, FL305DFY and FL305DPY. In Sahiwal cattle, mean ± SE of AA genotype for FL305DMY, FLTMY, FL305DFY, FL305DSNFY, FL305DPY were 1809.90 ± 15.7 kg, 2029.4 ± 15.6 kg, 99.90 ± 0.66 kg, 154.87 ± 0.17 kg and 44.81 ± 0.06 kg, respectively and for AB genotype were 1800.76 ± 9.48 kg, 1993.99 ± 9.42 kg, 100.54 ± 0.39 kg, 154.79 ± 0.10 kg, 43.99 ± 0.04 kg, respectively and for BB genotype were 1830.0 ± 14.10 kg, 2032.80 ± 14.0 kg, 100.24 ± 0.59 kg, 155.11 ± 0.15 kg, 42.98 ± 0.05 kg, respectively. Heterozygous AB genotype was found to be superior for, FL305DFY trait. AA genotype was significantly superior for FL305DPY traits whereas BB genotype was found to be superior for FL305DMY, FLTMY, FL305DSNFY. In Karan Fries cattle, the mean ± SE of AA genotype for FL305DMY, FLTMY, FL305DFY, FL305DSNFY, FL305DPY were 3442.17 ± 8.39 kg, 4461.93 ± 8.39 kg, 124.96 ± 7.20 kg, 277.35 ± 0.08 kg and 112.51 ± 0.08 kg, respectively and for AB genotype were 3572.69 ± 5.93 kg, 4592.45 ± 5.93 kg, 140.17 ± 5.09 kg, 278.60 ± 0.06 kg, 113.91 ± 0.05 kg, respectively and for BB genotypes were 3502.41 ± 9.19 kg, 4522.17 ± 9.19 kg, 136.91 ± 7.89 kg, 277.93 ± 0.09 kg, 113.19 ± 0.08 kg, respectively.

Use of the 16S–23S ribosomal genes spacer region for the molecular typing of sphingomonads

2008 ◽  
Vol 54 (8) ◽  
pp. 668-676 ◽  
Author(s):  
Sima Tokajian ◽  
Siba Al-Medawar ◽  
Fuad Hashwa
Keyword(s):  
Drinking Water ◽  
Molecular Typing ◽  
Opportunistic Infections ◽  
Its Region ◽  
Ribosomal Genes ◽  
Colony Morphology ◽  
Restriction Digestion ◽  
Banding Patterns ◽  
Hospital Acquired ◽  
Reference Strains

The ability of sphingomonads in drinking water to cause community- and hospital-acquired opportunistic infections has raised the need to establish reproducible identification assays. In this study, a total of 129 isolates recovered from drinking water with yellow- to orange-pigmented colonies were distributed among 10 biotypes on the basis of colony morphology. Polymorphisms, based on the amplification and restriction digestion of the intergenic transcribed spacer (ITS) region within the 10 assigned biotypes and 18 ATCC reference strains, were used to investigate the ability of this approach to differentiate closely related sphingomonads. ITS size, which ranged between 400 and 1100 bp, did not vary enough among the different genera. However, 16 distinct banding patterns within the ATCC reference strains and 9 within the 10 biotypes were obtained through ITS restriction digestion, and the majority of the tested biotypes produced patterns similar to those generated by the ATCC strains. To our knowledge, this study is not only the first comprehensive record of the size of the ITS region in sphingomonads, it is also the first study that describes the use of ITS restriction digestion to subtype those isolates.

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