scholarly journals Influence of host reproductive state onSphaerothecum destruensprevalence and infection level

Parasitology ◽  
2010 ◽  
Vol 138 (1) ◽  
pp. 26-34 ◽  
Author(s):  
D. ANDREOU ◽  
M. HUSSEY ◽  
S. W. GRIFFITHS ◽  
R. E. GOZLAN
Keyword(s):  
Detection Limit ◽  
Real Time ◽  
Genomic Dna ◽  
Adverse Impact ◽  
Intracellular Parasite ◽  
Reproductive State ◽  
Chain Reaction ◽  
Infection Level ◽  
Obligate Intracellular ◽  
Obligate Intracellular Parasite

SUMMARYSphaerothecum destruensis an obligate intracellular parasite with the potential to cause high mortalities and spawning inhibition in the endangered cyprinidLeucaspius delineatus. We investigated the influence ofL. delineatus’s reproductive state on the prevalence and infection level ofS. destruens. A novel real time quantitative polymerarse chain reaction (qPCR) was developed to determineS. destruens’ prevalence and infection level. These parameters were quantified and compared in reproductive and non-reproductiveL. delineatus. The detection limit of theS. destruensspecific qPCR was determined to be 1 pg of purifiedS. destruensgenomic DNA. Following cohabitation in the lab, reproductiveL. delineatushad a significantly higherS. destruensprevalence (P<0·05) and infection levels (P<0·01) compared to non-reproductiveL. delineatus. S. destruensprevalence was 19% (n=40) in non-reproductiveL. delineatusand 41% (n=32) in reproductiveL. delineatus. However, there was no difference inS. destruensprevalence in reproductive and non-reproductive fish under field conditions. Mean infection levels were 18 and 99 pgS. destruensDNA per 250 ngL. delineatusDNA for non-reproductive and reproductiveL. delineatusrespectively. The present work indicates thatS. destruensinfection inL. delineatuscan be influenced by the latter's reproductive state and provides further support for the potential adverse impact ofS. destruenson the conservation ofL. delineatuspopulations.

scholarly journals A selector of transcription initiation in the protozoan parasite Toxoplasma gondii.

1995 ◽  
Vol 15 (1) ◽  
pp. 87-93 ◽  
Author(s):  
D Soldati ◽  
J C Boothroyd
Keyword(s):  
Toxoplasma Gondii ◽  
Gene Transcription ◽  
Transcription Initiation ◽  
Protozoan Parasite ◽  
Intracellular Parasite ◽  
Transcription Start ◽  
Transcription Start Sites ◽  
Obligate Intracellular ◽  
Obligate Intracellular Parasite ◽  
Sag1 Gene

The recent development of an efficient transfection system for the apicomplexan Toxoplasma gondii allows a comprehensive dissection of the elements involved in gene transcription in this obligate intracellular parasite. We demonstrate here that for the SAG1 gene, a stretch of six repeated sequences in the region 35 to 190 bp upstream of the first of two transcription start sites is essential for efficient and accurate transcription initiation. This repeat element shows characteristics of a selector in determining the position of the transcription start sites.

scholarly journals Infected Dendritic Cells Facilitate Systemic Dissemination and Transplacental Passage of the Obligate Intracellular Parasite Neospora caninum in Mice

PLoS ONE ◽  
2012 ◽  
Vol 7 (3) ◽  
pp. e32123 ◽  
Author(s):  
Esther Collantes-Fernandez ◽  
Romanico B. G. Arrighi ◽  
Gema Álvarez-García ◽  
Jessica M. Weidner ◽  
Javier Regidor-Cerrillo ◽  
...  
Keyword(s):  
Dendritic Cells ◽  
Neospora Caninum ◽  
Intracellular Parasite ◽  
Transplacental Passage ◽  
Obligate Intracellular ◽  
Obligate Intracellular Parasite ◽  
Systemic Dissemination

Chlamydia isopodii sp. n., an Obligate Intracellular Parasite of Porcellio scaber

Pathobiology ◽  
1985 ◽  
Vol 53 (2) ◽  
pp. 115-120 ◽  
Author(s):  
Michael T. Shay ◽  
Annemarie Bettica ◽  
G.M. Vernon ◽  
E.R. Witkus
Keyword(s):  
Intracellular Parasite ◽  
Porcellio Scaber ◽  
Obligate Intracellular ◽  
Obligate Intracellular Parasite

Detection of the Escherichia coli pathogenic geneeaewith three real-time polymerase chain reaction methods

2007 ◽  
Vol 53 (3) ◽  
pp. 398-403 ◽  
Author(s):  
Joanne  Karen McCrea ◽  
Chenyi Liu ◽  
Lai-King Ng ◽  
Gehua Wang
Keyword(s):  
Escherichia Coli ◽  
Polymerase Chain Reaction ◽  
Detection Limit ◽  
Real Time ◽  
Turnaround Time ◽  
Sybr Green ◽  
Sybr Green I ◽  
Chain Reaction ◽  
Culture Techniques ◽  
Polymerase Chain

Several real-time polymerase chain reaction (PCR) methods are currently available to rapidly detect the presence of a specific DNA sequence. When used for detection of pathogenic organisms, the turnaround time for PCR-based methods is much lower than for traditional culture techniques. This study compared the sensitivity of three real-time PCR methods when detecting the Escherichia coli pathogenic gene eae to determine which method is most effective in identifying very low levels of the organism. The three methods were used to detect the eae gene over a range of DNA concentrations. The differences in sensitivity were statistically significant (p < 0.05), and SYBR Green I PCR was found to have the lowest detection limit of the three; LUX primers had the highest detection limit. Therefore, using a defined DNA concentration for detecting the eae gene, SYBR Green I is the best alternative.

scholarly journals Biotinylated Probe Isolation of Targeted Gene Region Improves Detection of T790M Epidermal Growth Factor Receptor Mutation via Peptide Nucleic Acid–Enriched Real-Time PCR

2011 ◽  
Vol 57 (5) ◽  
pp. 770-773 ◽  
Author(s):  
Jin Li ◽  
Pasi A Jänne ◽  
G Mike Makrigiorgos
Keyword(s):  
Epidermal Growth Factor Receptor ◽  
Detection Limit ◽  
Real Time ◽  
Genomic Dna ◽  
Mutant Allele ◽  
Growth Factor Receptor ◽  
Wild Type ◽  
T790m Mutation ◽  
Target Dna ◽  
Epidermal Growth

BACKGROUND The presence of the EGFR (epidermal growth factor receptor) T790M mutation in tumor tissue or body fluids from patients treated with EGFR tyrosine kinase inhibitors may indicate the onset of resistance to treatment. It is important to identify this mutation as early as possible so that treatment can be modified accordingly or potential side effects of further treatment can be avoided. This requirement calls for high detection sensitivity. Peptide nucleic acids (PNAs) are used as PCR clamps to inhibit amplification of wild-type DNA during PCR cycling, thereby enriching for rare mutations such as T790M. We describe a modification that improves the detection limit of PNA-clamp methods by at least 20-fold. METHODS We enriched the target by exposing genomic DNA to an EGFR exon 20–specific biotinylated oligonucleotide, followed by binding to streptavidin beads. We then prepared serial dilutions of the isolated target DNA containing the T790M mutation by mixing with wild-type DNA and then performed PNA clamp–based, real-time TaqMan PCR. For comparison, we performed PNA clamp–based PCR directly on genomic DNA. RESULTS Whereas the detection limit for PNA clamp–based PCR performed directly on genomic DNA is 1 mutant allele in 1000 wild-type alleles, conducting the assay with biotinylated oligonucleotide–enriched target DNA improved the detection limit to 1 mutant allele in 40 000 wild-type alleles. A possible explanation for the improvement in detection is that biotin-based target isolation efficiently eliminates wild-type DNA; therefore, fewer erroneous amplifications of wild-type DNA can occur early during the PCR. CONCLUSIONS Combining target molecule isolation via a biotinylated probe with PNA-enriched TaqMan real-time PCR provides a major improvement for detecting the EGFR T790M resistance mutation.

Acanthamoeba from human nasal mucosa infected with an obligate intracellular parasite

1994 ◽  
Vol 30 (1) ◽  
pp. 104-110 ◽  
Author(s):  
Rolf Michel ◽  
Bärbel Hauröder-Philippczyk ◽  
Karl-Dieter Müller ◽  
Iris Weishaar
Keyword(s):  
Nasal Mucosa ◽  
Intracellular Parasite ◽  
Human Nasal Mucosa ◽  
Obligate Intracellular ◽  
Obligate Intracellular Parasite
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