scholarly journals Determination of Types of Enterocytozoon bieneusiStrains Isolated from Patients with Intestinal Microsporidiosis

1998 ◽  
Vol 36 (7) ◽  
pp. 1882-1885 ◽  
Author(s):  
Olivier Liguory ◽  
Felicia David ◽  
Claudine Sarfati ◽  
Francis Derouin ◽  
Jean-Michel Molina
Keyword(s):  
Rflp Analysis ◽  
Its Region ◽  
Restriction Enzymes ◽  
Epidemiological Studies ◽  
Enterocytozoon Bieneusi ◽  
Type I ◽  
Pcr Rflp ◽  
Intestinal Microsporidiosis ◽  
Rflp Typing

To determine the types of Enterocytozoon bieneusistrains associated with intestinal microsporidiosis, we developed a rapid and efficient approach for typing parasites obtained from stool specimens by PCR-restriction fragment length polymorphism (PCR-RFLP). Typing was based on DNA polymorphism of the ribosomal DNA internal transcribed spacer (ITS) region of E. bieneusi. RFLPs generated with two restriction enzymes (NlaIII andFnu4HI) in PCR-amplified ITS products were used to classify strains into different lineages. This approach was successfully used to differentiate 78 strains that had been obtained from the stools of 65 patients with intestinal microsporidiosis. Among the 78 strains, we found four genetically unrelated lineages, showing the genetic diversity of E. bieneusi. Type I strains of E. bieneusi were found in a majority of the samples, accounting for 51 (78%) of the 65 microsporidiosis cases. In contrast, type II, III, and IV strains were found in only 8 (12%), 3 (5%), and 3 (5%) cases, respectively. All strains of E. bieneusi we have tested so far fall into one of four different lineages, and this study shows that human intestinal microsporidiosis is most often associated with type I strains. PCR-RFLP analysis of the ITS region of E. bieneusishould be useful for epidemiological studies.

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.

scholarly journals Genetic variation among pathogens causing "Helminthosporium" diseases of rice, maize and wheat

2002 ◽  
Vol 27 (6) ◽  
pp. 639-643 ◽  
Author(s):  
RITA C. B. WEIKERT-OLIVEIRA ◽  
M. APARECIDA DE RESENDE ◽  
HENRIQUE M. VALÉRIO ◽  
RACHEL B. CALIGIORNE ◽  
EDILSON PAIVA
Keyword(s):  
Triticum Aestivum ◽  
Genetic Variation ◽  
Climatic Factors ◽  
Rapd Analysis ◽  
Its Region ◽  
Restriction Enzymes ◽  
Fungal Species ◽  
High Similarity ◽  
Pcr Rflp ◽  
Upgma Phenogram

Twenty isolates of four fungal species, agents of "Helminthosporium" diseases in cereals, were collected from different regions: nine Bipolarisoryzae isolated from rice (Oryza sativa), seven B.sorokiniana from wheat (Triticum aestivum), two B. maydis, and two Exserohilumturcicum from maize (Zea mays). The strains were compared by PCR-RFLP and RAPD analysis. Size polymorphism among the isolates in the ITS region comprising the 5.8 S rDNA indicated genetic differences among the isolates, while a UPGMA phenogram constructed after the digestion of this region with restriction enzymes showed inter- and intra-specific polymorphism. The RAPD profiles indicated an expressive level of polymorphism among different species, compared with a low level of polymorphism among isolates of the same species. A UPGMA phenogram grouped the isolates according to the species and their host plant. RAPD profiles did not reveal polymorphism that directly correlated climatic factors with geographic source of the isolates of B. sorokiniana, and B. oryzae. Teleomorphic species revealed high similarity with their correspondent anamorphs.

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.

Steinernema apuliae sp. n. (Rhabditida: Steinernematidae): a new entomopathogenic nematode from southern Italy

Zootaxa ◽  
2004 ◽  
Vol 460 (1) ◽  
pp. 1 ◽  
Author(s):  
ORESTE TRIGGIANI ◽  
ZDENEK MRÁ»EK ◽  
ALEX REID
Keyword(s):  
First Generation ◽  
Southern Italy ◽  
Entomopathogenic Nematode ◽  
Rflp Analysis ◽  
Tail Length ◽  
Its Region ◽  
The Body ◽  
Steinernema Glaseri ◽  
Pcr Rflp ◽  
H 41

Steinernema apuliae sp. n. has been found in soil samples collected along a saltpan border habitat in southern Italy characterized by a salted silt soil. This species belongs to the long-IJ nematode group represented by Steinernema glaseri (Steiner, 1929) and Steinernema arenarium (Artyukhovsky, 1967) among others. However, it differs from these taxa in some morphometric values such as V%, H%. Females possess asymmetrical, oblique slit vulva, slant vagina and small flap in the vulval opening. These characteristics are more distinct in second generation females which is different from most other steinernematids; the vulva position is behind the mid-body about 57% to 61% of the body length. First-generation females have a conical-like tip bearing 2 to 3 papilla-like protuberances. Male mucron is absent in both generations. Lightly brown spicules have bluntly pointed tip and elongated manubrium. Third-stage infective juveniles are on average over 1000 m long; the position of the excretory pore is posterior (D% 66) and the hyaline layer is less than half the tail length (H% 41 42). Lateral fields are formed by 8 equally distributed ridges. S. apuliae differs from S. glaseri and S. arenarium and is separated by PCR-RFLP analysis of the ITS region. There were no positive cross-breedings among these species.

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.

Determination of flagellar types by PCR-RFLP analysis of enteropathogenic Escherichia coli (EPEC) and Shiga toxin-producing E. coli (STEC) strains isolated from animals in São Paulo, Brazil

2012 ◽  
Vol 92 (1) ◽  
pp. 18-23 ◽  
Author(s):  
Claudia de Oliveira Ayala ◽  
Ana Carolina Ramos Moreno ◽  
Marina Baquerizo Martinez ◽  
Ylanna Kelner Burgos ◽  
Antonio Fernando Pestana de Castro ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Shiga Toxin ◽  
Rflp Analysis ◽  
Sao Paulo ◽  
São Paulo ◽  
Enteropathogenic Escherichia Coli ◽  
E Coli ◽  
Pcr Rflp

PCR–RFLP analysis of mitochondrial DNA for identification of Rutilus rutilus caspicus populations on the southern coast of the Caspian Sea, Iran

2006 ◽  
Vol 86 (6) ◽  
pp. 1463-1467 ◽  
Author(s):  
Sohrab Rezvani ◽  
Amin Eimanifar ◽  
Reza Aghili ◽  
Faramarz Laloei
Keyword(s):  
Caspian Sea ◽  
Rflp Analysis ◽  
Restriction Enzymes ◽  
Rutilus Rutilus ◽  
Southern Coast ◽  
The Caspian Sea ◽  
Length Polymorphisms ◽  
Pcr Rflp ◽  
Nucleotide Divergence ◽  
Genetic Pools

Genetic analysis using restriction fragment length polymorphisms (RFLPs) of cytochrome b in mtDNA was made to clarify genetic variations among two Iranian Rutilus rutilus caspicus populations of commercial importance from the southern coast of the Caspian Sea. Polymorphism was detected using six restriction enzymes and a total of six composite haplotypes were identified. Four haplotypes were rare occurring only once in two regions (west and east of the southern Caspian Sea). Nucleotide and haplotype diversities were higher in the south-west region of the Caspian Sea (π=3.43%, h=23.3).The nucleotide divergence between the two populations was low (0.064%). The test for heterogeneity of composite haplotype frequencies gave no significant outcome for all samples (χ2=0.137, P≤0.05). The results indicate that significant attention should be paid to the genetic characterization of R. rutilus caspicus populations for conservation of their genetic pools and aquaculture policies at the coastlines of the Caspian Sea.

Characterization of Dryas octopetala ectomycorrhizas from limestone karst vegetation, western Ireland

2002 ◽  
Vol 80 (9) ◽  
pp. 970-982 ◽  
Author(s):  
Thomas J Harrington ◽  
Derek T Mitchell
Keyword(s):  
Rflp Analysis ◽  
Its Region ◽  
Fungal Species ◽  
Morphological Evidence ◽  
Soil Core ◽  
Cenococcum Geophilum ◽  
Dryas Octopetala ◽  
Pcr Rflp ◽  
Provisional Identification ◽  
Western Ireland

The principal ectomycorrhizas of Dryas octopetala L. from a treeless grass-heath in the Burren, western Ireland, were characterized using morphotyping and molecular methods (PCR-RFLP analysis of ITS-rDNA and sequencing of the ITS region). Twenty-one distinct morphotypes are described. Six of these (Cortinarius atrovirens Kalchbr., Cortinarius caesiocanescens (Mos.) Kühn. & Romagn., Cortinarius calochrous (Mos.) Nezd., Cortinarius odorifer Britz., Cortinarius mussivus Fr., and Tricholoma myomyces (Scop.) Quél.) were distinguished by tracing rhizomorph connections between mycorrhizas and basidiomes. The ectomycorrhizas of Cenococcum geophilum Fr., Craterellus lutescens (Pers.:Fr.) Fr., and Hebeloma sinapizans (Paulet:Fr.) Gill were identified based on molecular and morphological evidence. The ectomycorrhizas of Cortinarius brunneus (Pers.:Fr.) Fr., Cortinarius infractus (Pers.:Fr.) Fr., Hydnum repandum L., and Hebeloma circinans Quél. were distinguished provisionally, because they were consistently found in soil core samples containing basidiomes of a particular fungal species. The provisional identification of Lactarius sanguifluus (Paulet) Fr., and Russula delica Fr. ectomycorrhizas was also based on anatomical evidence, particularly the presence of lacticifers and cystidia, respectively. Six morphotypes could not be assigned to a specific fungal taxon and, therefore, were named "Dryadirhiza" + a characterizing epithet (D. aerea, D. cerina, D. fulgens, D. nigra, D. rugosa, and D. truncata). It is concluded that Dryas octopetala forms ectomycorrhizal associations in the Burren with woodland fungal species.Key words: ectomycorrhizas, Dryas octopetala, morphotyping, ITS-RFLP, mountain avens.

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 PHYLOGENETIC RELATIONSHIPS AMONGST 10 Durio SPECIES BASED ON PCR-RFLP ANALYSIS OF TWO CHLOROPLAST GENES

2013 ◽  
Vol 6 (1) ◽  
pp. 20 ◽  
Author(s):  
Panca J. Santoso ◽  
Ghizan B. Saleh ◽  
Norihan M. Saleh ◽  
Suhaimi Napis
Keyword(s):  
Phylogenetic Analysis ◽  
Phylogenetic Relationships ◽  
Rflp Analysis ◽  
Restriction Enzymes ◽  
Rbcl Gene ◽  
Research Station ◽  
Taxonomic Level ◽  
Pcr Rflp ◽  
Young Leaves ◽  
Reliable Marker

Twenty seven species of Durio have been identified in Sabah and Sarawak, Malaysia, but their relationships have not been studied. This study was conducted to analyse phylogenetic relationships amongst 10 Durio species in Malaysia using PCR-RFLP on two chloroplast DNA genes, i.e. ndhC-trnV and rbcL. DNAs were extracted from young leaves of 11 accessions from 10 Durio species collected from the Tenom Agriculture Research Station, Sabah, and University Agriculture Park, Universiti Putra Malaysia. Two pairs of oligonucleotide primers, N1-N2 and rbcL1-rbcL2, were used to flank the target regions ndhC-trnV and rbcL. Eight restriction enzymes, HindIII, BsuRI, PstI, TaqI, MspI, SmaI, BshNI, and EcoR130I, were used to digest the amplicons. Based on the results of PCR-RFLP on ndhC-trnV gene, the 10 Durio species were grouped into five distinct clusters, and the accessions generally showed high variations. However, based on the results of PCR-RFLP on the rbcL gene, the species were grouped into three distinct clusters, and generally showed low variations. This means that ndhC-trnV gene is more reliable for phylogenetic analysis in lower taxonomic level of Durio species or for diversity analysis, while rbcL gene is reliable marker for phylogenetic analysis at higher taxonomic level. PCR-RFLP on the ndhC-trnV and rbcL genes could therefore be considered as useful markers to phylogenetic analysis amongst Durio species. These finding might be used for further molecular marker assisted in Durio breeding program.

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