scholarly journals The Identification of Cryptosporidium Species by PCR-RFLP Analysis of the 18s rRNA Gene

2008 ◽  
Vol 12 ◽  
pp. e380-e381
Author(s):  
D. Dorostkar Moghaddam ◽  
M. Azami ◽  
R. Salehi ◽  
M. Salehi
Keyword(s):  
18S Rrna ◽  
Rflp Analysis ◽  
18S Rrna Gene ◽  
Cryptosporidium Species ◽  
Rrna Gene ◽  
Pcr Rflp

The identification of Cryptosporidium species in Isfahan, Iran by PCR-RFLP analysis of the 18S rRNA gene

2007 ◽  
Vol 41 (5) ◽  
pp. 851-856 ◽  
Author(s):  
M. Azami ◽  
D. D. Moghaddam ◽  
R. Salehi ◽  
M. Salehi
Keyword(s):  
18S Rrna ◽  
Rflp Analysis ◽  
18S Rrna Gene ◽  
Cryptosporidium Species ◽  
Rrna Gene ◽  
Pcr Rflp

scholarly journals Molecular Characterization ofCryptosporidiumspp. in Wild Rodents of Southwestern Iran Using 18s rRNA Gene Nested-PCR-RFLP and Sequencing Techniques

2016 ◽  
Vol 2016 ◽  
pp. 1-6 ◽  
Author(s):  
Jasem Saki ◽  
Masoud Foroutan-Rad ◽  
Reza Asadpouri
Keyword(s):  
Molecular Characterization ◽  
Nested Pcr ◽  
18S Rrna ◽  
Restriction Enzymes ◽  
18S Rrna Gene ◽  
Rrna Gene ◽  
Wild Rodents ◽  
Nested Polymerase Chain Reaction ◽  
Pcr Rflp ◽  
Southwest Of Iran

Background. Rodents could act as reservoir forCryptosporidiumspp. speciallyC. parvum, a zoonotic agent responsible for human infections. Since there is no information aboutCryptosporidiuminfection in rodents of Ahvaz city, southwest of Iran, hence, this survey was performed to determine the prevalence and molecular characterization ofCryptosporidiumspp. in this region.Materials and Methods. One hundred rodents were trapped from different regions of Ahvaz city. Intestine contents and fecal specimens of rodents were studied using both microscopy examination to identify oocyst and nested-polymerase chain reaction (PCR) technique for 18s rRNA gene detection. Eventually restriction fragment length polymorphism (RFLP) method usingSspIandVspIrestriction enzymes was carried out to genotype the species and then obtained results were sequenced.Results. Three out of 100 samples were diagnosed as positive and overall prevalence ofCryptosporidiumspp. was 3% using both modified Ziehl-Neelsen staining under light microscope and nested-PCR (830 bp) methods. Afterwards, PCR-RFLP was performed on positive samples andC. parvumpattern was identified. Finally PCR-RFLP findings were sequenced and presence ofC. parvumwas confirmed again.Conclusions. Our study showed rodents could be potential reservoir forC. parvum. So an integrated program for control and combat with them should be adopted and continued.

scholarly journals 6 Molecular typing of sand fly species by PCR-RFLP of 18S rRNA gene(Proceedings of the 63rd Annual Meeting of Western Region)

2009 ◽  
Vol 60 (2) ◽  
pp. 157
Author(s):  
H. Kato ◽  
Y. Terayama ◽  
A. Gomez Eduardo ◽  
H. Uezato ◽  
Calvopina Manuel ◽  
...  
Keyword(s):  
Annual Meeting ◽  
Molecular Typing ◽  
18S Rrna ◽  
18S Rrna Gene ◽  
Sand Fly ◽  
Rrna Gene ◽  
Western Region ◽  
Pcr Rflp

scholarly journals Molecular Screen of Trichoderma Isolates

1970 ◽  
Vol 17 ◽  
pp. 117-122
Author(s):  
Amna Ali ◽  
Rukhsana Bajwa ◽  
Nasir Mehmood ◽  
Rasheda Jabeen
Keyword(s):  
Gene Targeting ◽  
Genomic Dna ◽  
18S Rrna ◽  
Rflp Analysis ◽  
Stem Bark ◽  
18S Rrna Gene ◽  
Negative Control ◽  
Molecular Techniques ◽  
Rrna Gene ◽  
Trichoderma Species

Context: In the last few decades an ample array of molecular techniques has been introduced to obtain new disposition for the classification of Trichoderma species. Today the concern of scientists is either in the direction of gene targeting or ribotyping, the newest fingerprinting tool for genomic DNA that contain all or part of the genes coding for 18S rRNA in eukaryotes.Objectives: To take advantage of advanced molecular techniques for phylogenetic analysis of indigenous isolates of Trichoderma to comprehend our knowledge of this genus by supplementing the phenotypic identification.Materials and Methods:  Genomic DNA of twenty four isolates of Trichoderma species (T. harzianum, T. hamatum, T. koningii and T. pseudokoningii) were extracted by CTAB method and indicated band of ~15Kb on 0.8% agarose gel. Quality of DNA was determined by obtaining absorbance ratio (260/280) in the range of 1.7-1.9. Restriction fragment length polymorphism (RFLP) analyses were performed by using two restriction endonulease enzymes i.e., BamHI and HindIII. The BamHI represented results in the range of 500bp-750bp. 18S rRNA gene targeting was further carried out through optimization in ribotyping analysis.Results: The DNA bands of 24 isolates of Trichoderma species were compared with marker DNA bands and indicated the presence of genomic DNA intact band of ~15Kb. The ratio of absorbance 260/280nm (1.8-1.9 for pure DNA preparations) provided an estimate of the purity of the DNA RFLP analysis, along with the negative control of twenty four different isolates of Trichoderma species subjected to restriction using BamHI enzyme. The rRNA gene amplified band was observed at 600bp in the case of T. hamatum isolate (S. cumini stem bark, FCBP accession number 769) while in remaining isolates bands were in slightly smeared form. Furthermore, rRNA gene amplification conditions were optimized by altering different Tm and MgCl2 concentrations. Conclusion: The genomic DNA can serve as long term storage of information. Therefore advance molecular techniques can be used to study the variability in the genome of organism. RFLP are the initial steps for screening the genome of any organism.Key words: Trichoderma; DNA; RFLP; restriction endonuleases; 18S rRNA.DOI: 10.3329/jbs.v17i0.7117J. bio-sci. 17: 117-122, 2009

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