scholarly journals Unusual fatal avian polyomavirus infection in nestling cockatiels (Nymphicus hollandicus) detected by nested polymerase chain reaction

2008 ◽  
Vol 52 (No. 5) ◽  
pp. 193-201 ◽  
Author(s):  
O. Tomasek ◽  
O. Kubicek ◽  
V. Tukac
Keyword(s):  
Chain Reaction ◽  
Fatal Disease ◽  
Nested Polymerase Chain Reaction ◽  
Nymphicus Hollandicus ◽  
Chlamydophila Psittaci ◽  
Pcr Products ◽  
Polymerase Chain ◽  
Avian Polyomavirus ◽  
Feather Disease Virus ◽  
Nestling Mortality

High mortality of nestling cockatiels (<i>Nymphicus hollandicus</i>) was observed in one breeding flock in Slovakia. The nestling mortality affected 50% of all breeding pairs. In general, all the nestlings in affected nests died. Death occurred suddenly in 4- to 6-day-old birds, most of which had full crops. No feather disorders were diagnosed in this flock. Two dead nestlings were tested by nested PCR for the presence of avian polyomavirus (APV) and <i>Chlamydophila psittaci</i> and by single-round PCR for the presence of beak and feather disease virus (BFDV). After the breeding season ended, a breeding pair of cockatiels together with their young one and a fledgling budgerigar (<i>Melopsittacus undulatus</i>) were examined. No clinical alterations were observed in these birds. Haemorrhages in the proventriculus and irregular foci of yellow liver discoloration were found during necropsy in the young cockatiel and the fledgling budgerigar. Microscopy revealed liver necroses and acute haemolysis in the young cockatiel and confluent liver necroses and heart and kidney haemorrhages in the budgerigar. Two dead cockatiel nestlings, the young cockatiel and the fledgling budgerigar were tested positive for APV, while the cockatiel adults were negative. The presence of BFDV or <i>Chlamydophila psittaci</i> DNA was detected in none of the birds. The specificity of PCR was confirmed by the sequencing of PCR products amplified from the samples from the young cockatiel and the fledgling budgerigar. The sequences showed 99.6&minus;100% homology with the previously reported sequences. To our knowledge, this is the first report of APV infection which caused a fatal disease in parent-raised cockatiel nestlings and merely subclinical infection in budgerigar nestlings.

scholarly journals Specific Polymerase Chain Reaction Primers for the Detection of Plasmodiophora brassicae in Soil and Water

1999 ◽  
Vol 89 (5) ◽  
pp. 392-397 ◽  
Author(s):  
R. Faggian ◽  
S. R. Bulman ◽  
A. C. Lawrie ◽  
I. J. Porter
Keyword(s):  
Polymerase Chain Reaction ◽  
Plasmodiophora Brassicae ◽  
Chain Reaction ◽  
Nested Polymerase Chain Reaction ◽  
Simple Method ◽  
Soil P ◽  
Pcr Products ◽  
Polymerase Chain ◽  
Polymerase Chain Reaction Primers ◽  
Soil And Water

The development of specific oligonucleotide primers for Plasmodiophora brassicae has led to a nested polymerase chain reaction (PCR) detection method for P. brassicae in soil and water. Initially, the PCR was used to amplify a section of the rDNA repeat. The PCR products were sequenced and the data used to design primers that were directed at the ribosomal RNA genes and internal transcribed spacer regions. Specificity was tested against more than 40 common soil organisms, host plants, and spore suspension contaminants, as well as P. brassicae isolates from around Australia and the world. Sensitivity was determined to be 0.1 fentograms (fg; 10-15 g) for pure template and as low as 1,000 spores per g of potting mix. In soil, P. brassicae was detected in all soils where the inoculum was sufficient to result in clubroot symptoms. Also outlined is a simple method of DNA extraction from soil.

scholarly journals Detection of Canine Parvovirus in Baghdad city by PCR technique

2012 ◽  
Vol 36 (0E) ◽  
pp. 95-98
Author(s):  
Ahmed F. Ahmed
Keyword(s):  
Gel Electrophoresis ◽  
Canine Parvovirus ◽  
Chain Reaction ◽  
Viral Dna ◽  
Fatal Disease ◽  
The Third ◽  
Mutant Strains ◽  
Pcr Products ◽  
Polymerase Chain ◽  
Hemorrhagic Enteritis

Canine parvovirus 2 (CPV2) is a highly contagious and fatal disease of dogs, causingacute hemorrhagic enteritis and myocarditis. In this study different mutant strains of the viruswere characterized by polymerase chain reaction (PCR).The fecal samples from infected dogssuspected for CPV2 infection were collected in a suitable medium. The viral DNA from fecalsamples was extracted using specific kits, PCR were carried out with five different primer,pCPV-2ab and pCPV-2b, to distinguish the strain prevalent in field condition. The primerpCPV-2ab recognized both variant CPV-2a and CPV-2b, whereas the primer pCPV-2brecognized only the variant CPV-2b, using the third primer pCPV to recognize the residualbase pair, enabling the differentiation of CPV-2a variant from CPV-2b in field isolates. Thedifferent PCR products were further analyzed by using gel electrophoresis.

scholarly journals Molecular Identification ofEnterocytozoon bieneusiIsolates from Nigerian Children

2011 ◽  
Vol 2011 ◽  
pp. 1-2 ◽  
Author(s):  
Adekunle Bamidele Ayinmode ◽  
Oladele Teslim Ojuromi ◽  
Lihua Xiao
Keyword(s):  
Enterocytozoon Bieneusi ◽  
Chain Reaction ◽  
Nested Polymerase Chain Reaction ◽  
Nigerian Children ◽  
New Genotype ◽  
Pcr Products ◽  
Infection Source ◽  
Polymerase Chain ◽  
Immunocompetent Patients ◽  
High Diversity

A study was conducted to detect and identify enteric microsporidian species in 43 children from Oyo state, Nigeria. Using nested polymerase chain reaction, 9.3% of the children were identified as positive forEnterocytozoon bieneusi. DNA sequencing of the PCR products showed the presence of three known genotypes (two isolates of genotype D and one of genotype K) and one new genotype. This study suggests that either human or animal (or both) could be the infection source for the children, since identified genotypes D and K have been previously detected in both immunocompromised and immunocompetent patients and domestic animals. The identification of high diversity also suggests intensive transmission of microsporidiosis in the studied area.

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.

scholarly journals A Comparative Study of 5S rDNA Non-Transcribed Spacers in Elaeagnaceae Species

Plants ◽  
2020 ◽  
Vol 10 (1) ◽  
pp. 4
Author(s):  
Oleg S. Alexandrov ◽  
Olga V. Razumova ◽  
Gennady I. Karlov
Keyword(s):  
Polymerase Chain Reaction ◽  
Molecular Markers ◽  
Dna Markers ◽  
5S Rdna ◽  
Chain Reaction ◽  
Closely Related Species ◽  
Pcr Products ◽  
Polymerase Chain ◽  
Species Specific ◽  
Shepherdia Canadensis

5S rDNA is organized as a cluster of tandemly repeated monomers that consist of the conservative 120 bp coding part and non-transcribed spacers (NTSs) with different lengths and sequences among different species. The polymorphism in the 5S rDNA NTSs of closely related species is interesting for phylogenetic and evolutional investigations, as well as for the development of molecular markers. In this study, the 5S rDNA NTSs were amplified with universal 5S1/5S2 primers in some species of the Elaeagnaceae Adans. family. The polymerase chain reaction (PCR) products of five Elaeagnus species had similar lengths near 310 bp and were different from Shepherdia canadensis (L.) Nutt. and Sh. argentea (Pusch.) Nutt. samples (260 bp and 215 bp, respectively). The PCR products were cloned and sequenced. An analysis of the sequences revealed that intraspecific levels of NTS identity are high (approximately 95–96%) and similar in the Elaeagnus L. species. In Sh. argentea, this level was slightly lower due to the differences in the poly-T region. Moreover, the intergeneric and intervarietal NTS identity levels were studied and compared. Significant differences between species (except E. multiflora Thunb. and E. umbellata Thunb.) and genera were found. Herein, a range of the NTS features is discussed. This study is another step in the investigation of the molecular evolution of Elaeagnaceae and may be useful for the development of species-specific DNA markers in this family.

Detection of hepatitis “C” virus in formalin-fixed liver tissue by nested polymerase chain reaction

1992 ◽  
Vol 37 (4) ◽  
pp. 310-314 ◽  
Author(s):  
Richard Sallie ◽  
Anne Rayner ◽  
Bernard Portmann ◽  
A. L. W. F. Eddleston ◽  
Roger Williams
Keyword(s):  
Polymerase Chain Reaction ◽  
Hepatitis C Virus ◽  
Hepatitis C ◽  
Liver Tissue ◽  
Chain Reaction ◽  
Nested Polymerase Chain Reaction ◽  
Polymerase Chain ◽  
Formalin Fixed

The fate of a genetically modifiedPseudomonasstrain and its transgene during the composting of poultry manure

2004 ◽  
Vol 50 (6) ◽  
pp. 415-421 ◽  
Author(s):  
J Guan ◽  
J L Spencer ◽  
M Sampath ◽  
J Devenish
Keyword(s):  
Polymerase Chain Reaction ◽  
Incubation Temperature ◽  
Genetically Modified ◽  
Chicken Manure ◽  
Detection Sensitivity ◽  
Plate Count ◽  
Chain Reaction ◽  
Nested Polymerase Chain Reaction ◽  
Soil Microcosms ◽  
Polymerase Chain

The fate of the genetically modified (GM) Pseudomonas chlororaphis strain 3732 RN-L11 and its transgene (lacZ insert) during composting of chicken manure was studied using plate count and nested polymerase chain reaction (PCR) methods. The detection sensitivity of the nested PCR method was 165 copies of the modified gene per gram of moist compost or soil. Compost microcosms consisted of a 100-g mixture of chicken manure and peat, whereas soil microcosms were 100-g samples of sandy clay loam. Each microcosm was inoculated with 4 × 1010CFU of P. chlororaphis RN-L11. In controlled temperature studies, neither P. chlororaphis RN-L11 nor its transgene could be detected in compost microcosms after incubation temperature was elevated to 45 °C or above for one or more days. In contrast, in the compost microcosms incubated at 23 °C, the target organism was not detected by the plate count method after 6 days, but its transgene was detectable for at least 45 days. In compost bins, the target organism was not recovered from compost microcosms or soil microcosms at different levels in the bins for 29 days. However, the transgene was detected in 8 of the 9 soil microcosms and in only 1 of the 9 compost microcosms. The compost microcosm in which transgene was detected was at the lower level of the bin where temperatures remained below 45 °C. The findings indicated that composting of organic wastes could be used to reduce or degrade heat sensitive GM microorganisms and their transgenes.Key words: composting, genetically modified Pseudomonas strain, transgene, polymerase chain reaction.

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