A molecular phylogeographic study based on DNA sequences from individual metacercariae ofParagonimus mexicanusfrom Guatemala and Ecuador

2003 ◽  
Vol 77 (1) ◽  
pp. 33-38 ◽  
Author(s):  
M. Iwagami ◽  
C. Monroy ◽  
M.A. Rosas ◽  
M.R. Pinto ◽  
A.G. Guevara ◽  
...  
Keyword(s):  
Dna Sequences ◽  
Internal Transcribed Spacer ◽  
Distinct Species ◽  
Genetic Distances ◽  
Ribosomal Gene ◽  
Chain Reaction ◽  
Geographic Origins ◽  
Polymerase Chain ◽  
Second Internal Transcribed Spacer ◽  
Phylogeographic Study

AbstractA molecular phylogeographic study ofParagonimus mexicanuscollected from Guatemala and Ecuador was performed. Genomic DNA was extracted from individual metacercariae, and two gene regions (partial mitochondrial cytochromecoxidase subunit 1 (CO1) and the second internal transcribed spacer of the nuclear ribosomal gene repeat (ITS2)) were amplified by the polymerase chain reaction (PCR). Sequences segregated in a phylogenetic tree according to their geographic origins. ITS2 sequences from Ecuador and Guatemala differed at only one site. Pairwise distances among CO1 sequences within a country were always lower than between countries. Nevertheless, genetic distances between countries were less than between geographical forms ofP. westermanithat have been suggested to be distinct species. This result suggests that populations from Guatemala and Ecuador are genetically differentiated perhaps at the level of subspecies.

scholarly journals Geographical genetic structure within the human lung fluke, Paragonimus westermani, detected from DNA sequences

Parasitology ◽  
1997 ◽  
Vol 115 (4) ◽  
pp. 411-417 ◽  
Author(s):  
D. BLAIR ◽  
T. AGATSUMA ◽  
T. WATANOBE ◽  
M. OKAMOTO ◽  
A. ITO
Keyword(s):  
Dna Sequences ◽  
Distinct Species ◽  
Genetic Distances ◽  
The Philippines ◽  
Peninsular Malaysia ◽  
Ribosomal Gene ◽  
Paragonimus Westermani ◽  
Lung Fluke ◽  
Coi Sequences ◽  
Second Internal Transcribed Spacer

Nucleotide sequences were obtained for the second internal transcribed spacer of the ribosomal gene repeat and for part of the mitochondrial cytochrome c oxidase subunit I gene from geographical isolates of Paragonimus westermani from Japan, China, Korea, Taiwan, the Philippines, peninsular Malaysia and Thailand. Sequences were obtained from several other species of Paragonimus for comparative purposes. Two groups were recognized within P. westermani: an NE group (China, Japan, Korea, Taiwan) which was relatively uniform and included both diploid and triploid forms, and a southern group (Malaysia, Thailand, Philippines), members of which were genetically distant from one another. According to both ITS2 and COI data, genetic distances among P. westermani isolates equalled or exceeded those between some distinct species of Paragonimus. The ITS2 sequences were conserved relative to COI sequences. Substitutions among the latter may be approaching saturation within the genus Paragonimus.

scholarly journals Molecular confirmation of Xiphinema italiae Meyl, 1953 (Nematoda: Longidoridae) from the Slovak Republic

2009 ◽  
Vol 46 (2) ◽  
pp. 131-134 ◽  
Author(s):  
S. Kumari ◽  
M. Lišková
Keyword(s):  
Polymerase Chain Reaction ◽  
Molecular Markers ◽  
Cytochrome C Oxidase ◽  
Internal Transcribed Spacer ◽  
Slovak Republic ◽  
Chain Reaction ◽  
Polymerase Chain Reaction Protocol ◽  
Specific Identification ◽  
Polymerase Chain ◽  
Second Internal Transcribed Spacer

AbstractIdentification of the nematode Xiphinema italiae relies mainly on time-consuming morphological and morphometrical studies. A polymerase chain reaction protocol has been used for the reliable and specific identification of X. italiae. Moreover, four independently evolving molecular markers (cox1- cytochrome c oxidase subunit 1; ITS2-second internal transcribed spacer; 18S gene and D2/D3 expansion segments of 28S gene) were amplified and sequenced in both directions.

Using a Polymerase Chain Reaction as a Complementary Test to Improve the Detection of Dermatophyte Fungus in Nails

2014 ◽  
Vol 104 (3) ◽  
pp. 233-237 ◽  
Author(s):  
María José Iglesias Sánchez ◽  
Ana María Pérez Pico ◽  
Félix Marcos Tejedor ◽  
María Jesús Iglesias Sánchez ◽  
Raquel Mayordomo Acevedo
Keyword(s):  
Polymerase Chain Reaction ◽  
Dna Sequences ◽  
Classical Method ◽  
Clinical Manifestations ◽  
Culture Method ◽  
Clinical Samples ◽  
Chain Reaction ◽  
Nested Polymerase Chain Reaction ◽  
Traditional Procedure ◽  
Polymerase Chain

Background Dermatomycoses are a group of pathologic abnormalities frequently seen in clinical practice, and their prevalence has increased in recent decades. Diagnostic confirmation of mycotic infection in nails is essential because there are several pathologic conditions with similar clinical manifestations. The classical method for confirming the presence of fungus in nail is microbiological culture and the identification of morphological structures by microscopy. Methods We devised a nested polymerase chain reaction (PCR) that amplifies specific DNA sequences of dermatophyte fungus that is notably faster than the 3 to 4 weeks that the traditional procedure takes. We compared this new technique and the conventional plate culture method in 225 nail samples. The results were subjected to statistical analysis. Results We found concordance in 78.2% of the samples analyzed by the two methods and increased sensitivity when simultaneously using the two methods to analyze clinical samples. Now we can confirm the presence of dermatophyte fungus in most of the positive samples in just 24 hours, and we have to wait for the result of culture only in negative PCR cases. Conclusions Although this PCR cannot, at present, substitute for the traditional culture method in the detection of dermatophyte infection of the nails, it can be used as a complementary technique because its main advantage lies in the significant reduction of time used for diagnosis, in addition to higher sensitivity.

Polymerase chain reaction based mapping of rye involving repeated DNA sequences

Genome ◽  
1992 ◽  
Vol 35 (4) ◽  
pp. 621-626 ◽  
Author(s):  
Peter M. Rogowsky ◽  
Ken W. Shepherd ◽  
Peter Langridge
Keyword(s):  
Polymerase Chain Reaction ◽  
Dna Sequences ◽  
Chromosome 1 ◽  
Repeated Dna ◽  
Chain Reaction ◽  
Pcr Marker ◽  
Banding Patterns ◽  
Repeated Dna Sequences ◽  
Cereal Rye ◽  
Polymerase Chain

A novel type of polymerase chain reaction (PCR) marker was developed for the mapping of cereal rye (Secale cereale). Primer pairs were synthesized targeting the insertion sites of three individual copies of the R173 family of rye specific repeated DNA sequences. While one primer was derived from a sequence within the respective R173 element, the second primer corresponded to a flanking region. The complex banding patterns obtained in rye allowed not only the mapping of the three R173 elements to certain chromosome regions of 1RS (the short arm of rye chromosome 1) but also the mapping of an additional 3–10 easily identifiable bands per primer pair to other rye chromosomes. Linkage mapping of a polymorphic 1R band derived from three rye cultivars demonstrated the presence of nonallelic, dominant markers in two independent crosses. Because of the high copy number of the R173 family (15 000 copies per diploid rye genome), its dispersion over the entire length of all chromosomes and the high number of markers obtained per primer pair, PCR markers based on the R173 family provide an almost unlimited source for well-spaced markers in rye mapping.Key words: polymerase chain reaction, mapping, repetitive DNA sequences, wheat, rye.

Genetic diversity and phylogeny analysis of Azolla based on DNA amplification by arbitrary primers

Genome ◽  
1993 ◽  
Vol 36 (4) ◽  
pp. 686-693 ◽  
Author(s):  
Benoit Van Coppenolle ◽  
Iwao Watanabe ◽  
Charles Van Hove ◽  
Gerard Second ◽  
Ning Huang ◽  
...  
Keyword(s):  
Principal Component Analysis ◽  
Polymerase Chain Reaction ◽  
Principal Component ◽  
Dna Amplification ◽  
Component Analysis ◽  
Genetic Distances ◽  
Group Method ◽  
Chain Reaction ◽  
Arbitrary Primers ◽  
Polymerase Chain

The polymerase chain reaction was used to amplify random sequences of DNA from 25 accessions of Azolla to evaluate the usefulness of this technique for identification and phylogenetic analysis of this aquatic fern. Accessions were selected to represent all known species within the genus Azolla and to encompass the worldwide distribution of the fern. Primers of 10 nucleotides with 70% G + C content were used to generate randomly amplified polymorphic DNA from the symbiotic Azolla–Anabaena complex. Twenty-two primers were used and each primer gave 4–10 bands of different molecular weights for each accession. Bands were scored as present or absent for each accession and variation among accessions was quantified using Nei's genetic distances. A dendrogram summarizing phenetic relationships among the 25 accessions was generated using the unweighted pair-group method with arithmetic mean. Principal component analysis was also used to evaluate genetic similarities. Three distinct groups were identified: group 1 contains five species, group 2 contains the pinnata species, and group 3 contains the nilotica species. The analysis demonstrates that the major groups of Azolla species can be easily distinguished from one an other and, in addition, that closely related accessions within species can be identified. We further found that using 10 primers, a phylogeny that is essentially the same as that derived from 22 primers can be constructed. Our results suggest that total DNA extracted from the Azolla–Anabaena symbionts is useful for classification and phylogenetic studies of Azolla.Key words: Azolla–Anabaena symbiosis, genetic distances, polymerase chain reaction, principal component analysis.

scholarly journals Molecular typing of bacteria of the genus Asaia in malaria vector Anopheles arabiensis Patton, 1905

2012 ◽  
Vol 44 (2) ◽  
pp. 7 ◽  
Author(s):  
S. Epis ◽  
M. Montagna ◽  
F. Comandatore ◽  
C. Damiani ◽  
A. Diabaté ◽  
...  
Keyword(s):  
Molecular Typing ◽  
Internal Transcribed Spacer ◽  
Anopheles Arabiensis ◽  
Acetic Acid Bacterium ◽  
Sub Saharan Africa ◽  
Randomly Amplified Polymorphic Dna ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Sub Saharan ◽  
Dna Pcr

The acetic acid bacterium <em>Asaia</em> spp. was successfully detected in <em>Anopheles arabiensis</em> Patton, 1905, one of the major vector of human malaria in Sub-Saharan Africa. A collection of 45 <em>Asaia</em> isolates in cellfree media was established from 20 individuals collected from the field in Burkina Faso. 16S rRNA universal polymerase chain reaction (PCR) and specific qPCR, for the detection of <em>Asaia</em> spp. were performed in order to reveal the presence of different bacterial taxa associated with this insect. The isolates were typed by internal transcribed spacer-PCR, BOX-PCR, and randomly amplified polymorphic DNA-PCR, proved the presence of different <em>Asaia</em> in <em>A. arabiensis</em>.

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