scholarly journals PositivePneumocystis jiroveciiSputum PCR Results with Negative Bronchoscopic PCR Results in Suspected Pneumocystis Pneumonia

2018 ◽  
Vol 2018 ◽  
pp. 1-5 ◽  
Author(s):  
Kelly Pennington ◽  
John Wilson ◽  
Andrew H. Limper ◽  
Patricio Escalante
Keyword(s):  
Polymerase Chain Reaction ◽  
Chart Review ◽  
Time Frame ◽  
Pneumocystis Pneumonia ◽  
Negative Sample ◽  
Sample Collection ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Immunocompromised State ◽  
Tracheal Secretion

Introduction. The diagnostic standard forPneumocystis jiroveciipneumonia (PCP) is direct microscopic identification; however, in recent years, polymerase chain reaction (PCR) from bronchoalveolar lavage (BAL) samples to detectPneumocystisnucleic acids has proven to be more sensitive and specific. Sputum samples have been presumed inferior to bronchoscopic samples secondary to variability and adequacy of sample collection. We observed several cases of positive sputum PCP-PCR results with negative PCP-PCR BAL results. The aim of the current study was to further characterize the clinical setting and outcomes in patients with positive sputum PCP-PCR samples and negative BAL PCP-PCR samples.Methods. We identified all patients who underwentP. jirovecii-PCR testing at Mayo Clinic between 2011 and 2016. Patients with a positive sputum and negative BAL sample collected within a 14-day time frame were identified and underwent further chart review for demographics, immunocompromised state, and clinical outcome.Results. From 2011 to 2016, 4431 respiratory samples from 3021 unique patients were tested for the presence ofP. jiroveciiby PCR. Fifty-five samples (1.2% of all samples collected) belonging to 24 unique patients (0.79% of patients tested) were identified as having a positive and negative sample collected within 14 days. Of these 24 patients, 10 (46%) patients had a positive sputum or tracheal secretion sample with negative BAL or bronchial washings. Out of these 10 patients, 8 were immunocompromised and 9 underwent treatment for PCP with 6 patients improving.Conclusion. Our results suggest that discordantP. jirovecii-PCR testing results from sputum and bronchoscopic specimens are an infrequent occurrence. Patients with positiveP. jirovecii-PCR sputum/tracheal secretion samples and negative bronchoscopic samples appear to be clinically infected and respond to PCP treatment. SputumP. jirovecii-PCR testing may be a viable alternative to invasive testing.

scholarly journals Accuracy of quantitative polymerase chain reaction in samples of frozen and paraffin-embedded healthy skin for the diagnosis of canine visceral leishmaniasis

2017 ◽  
Vol 69 (6) ◽  
pp. 1443-1450 ◽  
Author(s):  
M.P. Campos ◽  
M.F. Madeira ◽  
D.A. Silva ◽  
M.S. Solcà ◽  
O.M. Espíndola ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Quantitative Polymerase Chain Reaction ◽  
Healthy Skin ◽  
Sample Collection ◽  
Chain Reaction ◽  
Dna Recovery ◽  
Ffpe Samples ◽  
Fresh Frozen ◽  
Polymerase Chain ◽  
Commercial Kits

ABSTRACT The purpose of the present work was to evaluate the accuracy of quantitative polymerase chain reaction (qPCR) performed on samples of fresh frozen tissue (FT) and formalin-fixed, paraffin-embedded (FFPE) healthy skin. This is a validation study conducted with samples from 46 dogs from an endemic area in Brazil. After sample collection, DNA extractions were conducted using commercial kits and qPCR was oriented to kinetoplast DNA (kDNA) targets of the Leishmania infantum species. The results obtained for the FFPE samples showed 63.6% sensitivity and 77.1% specificity, whereas those obtained for the FT samples showed 100% and 48.6%, respectively. Poor agreement was observed for the results of the qPCR technique with FT and FFPE samples. Our results suggest freezing as the most suitable conservation method for the formation of sample databases considering DNA recovery

scholarly journals Psittacine beak and feather disease virus in budgerigars and ring-neck parakeets in South Africa

2004 ◽  
Vol 71 (1) ◽  
Author(s):  
J. Albertyn ◽  
K.M. Tajbhai ◽  
R.R. Bragg
Keyword(s):  
South Africa ◽  
Polymerase Chain Reaction ◽  
Genetic Variation ◽  
Disease Virus ◽  
Common Disease ◽  
Sample Collection ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Feather Disease Virus ◽  
Viral Isolates

Psittacine beak and feather disease (PBFD) is a common disease of the psittacine species and is caused by the psittacine beak and feather disease virus (PBFDV). In this study the occurrence of the disease in ring-neck parakeets and budgerigars in South Africa suffering from feathering problems, using polymerase chain reaction as a diagnostic test was investigated. The genetic variation between viral isolates was also studied. Results indicate that PBFDV can be attributed to being the cause of feathering problems in some of the ring-neck parakeets and budgerigars in South Africa. Genetic variation of isolates occurs between species and individuals. A cheap and easy to use method of blood sample collection on filter paper for diagnostic purposes was also evaluated. It proved to be less stressful to the birds and did not inhibit further processes.

Diagnosis of pneumocystis pneumonia in a 2-year-old King Charles Cavalier Spaniel using the polymerase chain reaction

2018 ◽  
Vol 47 (1) ◽  
pp. 146-149 ◽  
Author(s):  
Ayeley A. K. Okine ◽  
Seth Chapman ◽  
Roger A. Hostutler ◽  
Robert Livingston
Keyword(s):  
Polymerase Chain Reaction ◽  
Pneumocystis Pneumonia ◽  
Chain Reaction ◽  
Polymerase Chain

scholarly journals Simplified Method of Detection of Dialister invisus and Olsenella uli in Oral Cavity Samples by Polymerase Chain Reaction

2017 ◽  
Vol 8 (1-2) ◽  
pp. 47-52 ◽  
Author(s):  
Manohar S. Kugaji ◽  
Kishore G. Bhat ◽  
Vinayak M. Joshi ◽  
Madhu Pujar ◽  
Pratik T. Mavani
Keyword(s):  
Polymerase Chain Reaction ◽  
Bacterial Species ◽  
Universal Primers ◽  
Sample Collection ◽  
Chain Reaction ◽  
Infection Group ◽  
Endodontic Infection ◽  
Predominant Component ◽  
Endodontic Infections ◽  
Polymerase Chain

Aims and Objectives: The oral microbial flora is highly complex and diverse with obligate anaerobic bacteria as the predominant component. Most of these are not yet cultivated/difficult to cultivate due to technical limitations. In this study, we aim to detect two novel oral bacterial species Dialister invisus and Olsenella uli by simplified and economical procedure of polymerase chain reaction (PCR) and study their association with primary and persistent endodontic infections. Material and Methods: The study involved 60 patients that included 30 patients of primary endodontic infections and 30 with persistent endodontic infections. The sample collection from the root canal was performed by universally accepted protocol by using sterile paper points. The deoxyribonucleic acid (DNA) extraction was done, followed by PCR with species specific primers. We made several changes to the protocol mentioned by original authors. We adopted a one-step protocol for amplification of bacterial DNA, omitting the 16SrDNA amplification step with universal primers. Results: It was seen that 7 (23.3 %) samples in primary endodontic infection group and 24 (80 %) samples in persistent endodontic infection group were positive for D. invisus. On the other hand, 11 (36.6 %) patients of primary endodontic infection showed positivity for O. uli in comparison to 9 (30 %) of persistent endodontic infection. Conclusion: The results from the present study showed efficient amplification of both O. uli and D. invisus in a single-step PCR. Hence, we conclude that the modified protocol used here with taq polymerase enzyme offers a faster and cheaper alternative to nested PCR without compromising the quality of amplification process.

Low Sensitivity of a Nested Polymerase Chain Reaction in Oropharyngeal Washings for the Diagnosis of Pneumocystis Pneumonia in HIV-Infected Patients

CHEST Journal ◽  
2005 ◽  
Vol 128 (1) ◽  
pp. 167-171 ◽  
Author(s):  
Kennedy Nyamande ◽  
Umesh G. Lalloo ◽  
Dennis York ◽  
Mogambal Naidoo ◽  
Elvis M. Irusen ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Pneumocystis Pneumonia ◽  
Chain Reaction ◽  
Nested Polymerase Chain Reaction ◽  
Polymerase Chain ◽  
Low Sensitivity

scholarly journals Detection of Mycoplasma agalactiae by Polymerase Chain Reaction in Jordanian Sheep and Goat Herds

2007 ◽  
Vol 76 (1) ◽  
pp. 71-77 ◽  
Author(s):  
D. Zendulková ◽  
A. Madanat ◽  
P. Lány ◽  
K. Rosenbergová ◽  
Z. Pospíšil
Keyword(s):  
Polymerase Chain Reaction ◽  
Clinical Signs ◽  
Mycoplasma Species ◽  
Sample Collection ◽  
Chain Reaction ◽  
Mycoplasma Agalactiae ◽  
Sheep And Goats ◽  
Contagious Agalactia ◽  
Polymerase Chain ◽  
First Time

The aim of the study was to ascertain whether sheep and goats from selected Jordanian herds were infected with Mycoplasma agalactiae, the most common aetiological agent of contagious agalactia of sheep and goats. All examined animals showed clinical signs of disease at the time of sample collection. The group included 35 animals, 15 sheep and 20 goats. For microbiological examination, a total of 107 swabs were taken from conjunctival, nasal, vaginal or preputial mucosae and from the external auditory canal. Identification of the species isolated was carried out by a polymerase chain reaction. Of the 35 animals, 21 (4 sheep and 17 goats) tested positive for Mycoplasma agalactiae. These results confirmed our assumption that this mycoplasma species is present in Jordanian herds and, for the first time, provided evidence that contagious agalactia of sheep and goats occurs in Jordan.

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