scholarly journals A Qualitative Seminested PCR Assay as an Alternative to Urine Cytology for BK Polyomavirus Screening after Renal Transplantation

Intervirology ◽  
2013 ◽  
Vol 56 (4) ◽  
pp. 249-252 ◽  
Author(s):  
Ana Carolina Jonard Zalona ◽  
Rafael Brandão Varella ◽  
Cristina Maeda Takiya ◽  
Renato Torres Goncalves ◽  
Mariano Gustavo Zalis ◽  
...  
Keyword(s):  
Renal Transplantation ◽  
Urine Cytology ◽  
Pcr Assay ◽  
Bk Polyomavirus ◽  
Seminested Pcr

scholarly journals Reactivation of BK Polyomavirus in Urine Cytology Is Not Associated with Urothelial Cell Carcinoma

Viruses ◽  
2020 ◽  
Vol 12 (12) ◽  
pp. 1412
Author(s):  
Faisal Klufah ◽  
Ghalib Mobaraki ◽  
Axel zur Hausen ◽  
Iryna V. Samarska
Keyword(s):  
Cell Carcinoma ◽  
Urine Cytology ◽  
Urothelial Cell ◽  
The Other ◽  
Urothelial Cell Carcinoma ◽  
Tissue Samples ◽  
Bk Polyomavirus ◽  
Ffpe Tissues ◽  
Immunosuppressed Patients ◽  
History Of

BK polyomavirus (BKPyV) has been associated with some high-grade and special urothelial cell carcinoma (UCC) subtypes in immunosuppressed patients. Here, we evaluated the relationship of BKPyV-positive urine cytology specimens (UCS) with UCC. A large single-institution database was retrospectively searched for UCS positive for decoy cells, suggesting BKPyV infection. These were tested for the presence of BKPyV by PCR and immunohistochemistry (IHC) in urine sediments and formalin-fixed paraffin-embedded (FFPE) tissue samples of UCC. Decoy cells were reported in 30 patients out of the database with 22.867 UCS. Of these 30 patients, 16 (53.3%) had no history of UCC. Six patients out of these 16 had a history of transplantation, 4 had a history of severe chronic medical conditions, and 6 had no chronic disease. The other fourteen patients were diagnosed with either in situ or invasive UCC of the urinary bladder (14/30; 46.6%) prior to the detection of decoy cells in the urine. Nine of these UCC patients received intravesical treatment (BCG or mitomycin) after the first presentation with UCC. However, the clinical data on the treatment of the other five UCC patients was lacking. IHC identified BKPyV-positivity in the urine samples of non-UCC and UCC patients, while no BKPyV positivity was found in FFPE tissues of primary UCCs and metastases. In addition, BKPyV-PCR results revealed the presence of BKPyV DNA in the urine of the UCC cases, yet none in the UCC tissues itself. These data strongly indicate that BKPyV reactivation is not restricted to immunosuppression. It can be found in UCS of the immunocompetent patients and may be related to the intravesical BCG or mitomycin treatment of the UCC patients.

scholarly journals Detection of Small Numbers of Campylobacter jejuni and Campylobacter coli Cells in Environmental Water, Sewage, and Food Samples by a Seminested PCR Assay

1999 ◽  
Vol 65 (4) ◽  
pp. 1636-1643 ◽  
Author(s):  
Astrid S. Waage ◽  
Traute Vardund ◽  
Vidar Lund ◽  
Georg Kapperud
Keyword(s):  
Campylobacter Jejuni ◽  
Simple Procedure ◽  
Environmental Water ◽  
Food Samples ◽  
Proteinase K ◽  
Coliform Bacteria ◽  
Intergenic Sequence ◽  
Campylobacter Coli ◽  
Pcr Assay ◽  
Seminested Pcr

ABSTRACT A rapid and sensitive assay was developed for detection of small numbers of Campylobacter jejuni and Campylobacter coli cells in environmental water, sewage, and food samples. Water and sewage samples were filtered, and the filters were enriched overnight in a nonselective medium. The enrichment cultures were prepared for PCR by a rapid and simple procedure consisting of centrifugation, proteinase K treatment, and boiling. A seminested PCR based on specific amplification of the intergenic sequence between the two Campylobacter flagellin genes, flaA andflaB, was performed, and the PCR products were visualized by agarose gel electrophoresis. The assay allowed us to detect 3 to 15 CFU of C. jejuni per 100 ml in water samples containing a background flora consisting of up to 8,700 heterotrophic organisms per ml and 10,000 CFU of coliform bacteria per 100 ml. Dilution of the enriched cultures 1:10 with sterile broth prior to the PCR was sometimes necessary to obtain positive results. The assay was also conducted with food samples analyzed with or without overnight enrichment. As few as ≤3 CFU per g of food could be detected with samples subjected to overnight enrichment, while variable results were obtained for samples analyzed without prior enrichment. This rapid and sensitive nested PCR assay provides a useful tool for specific detection of C. jejuni or C. coli in drinking water, as well as environmental water, sewage, and food samples containing high levels of background organisms.

Development and evaluation of a one-tube seminested PCR assay for the detection and identification of Penicillium marneffei

Mycoses ◽  
2003 ◽  
Vol 46 (11-12) ◽  
pp. 447-454 ◽  
Author(s):  
C. Prariyachatigul ◽  
A. Chaiprasert ◽  
K. Geenkajorn ◽  
R. Kappe ◽  
C. Chuchottaworn ◽  
...  
Keyword(s):  
Penicillium Marneffei ◽  
Pcr Assay ◽  
Detection And Identification ◽  
Seminested Pcr

scholarly journals Urine cytology screening for polyoma virus infection following renal transplantation: the Oxford experience

2006 ◽  
Vol 60 (8) ◽  
pp. 927-930 ◽  
Author(s):  
T. P Thamboo ◽  
K. J M Jeffery ◽  
P. J Friend ◽  
G. D H Turner ◽  
I. S D Roberts
Keyword(s):  
Renal Transplantation ◽  
Virus Infection ◽  
Polyoma Virus ◽  
Urine Cytology ◽  
Cytology Screening

scholarly journals Application of pbp1A PCR in Identification of Penicillin-Resistant Streptococcus pneumoniae

1999 ◽  
Vol 37 (3) ◽  
pp. 628-632 ◽  
Author(s):  
Mignon du Plessis ◽  
Anthony M. Smith ◽  
Keith P. Klugman
Keyword(s):  
Streptococcus Pneumoniae ◽  
Clinical Isolates ◽  
Penicillin Resistance ◽  
Binding Region ◽  
Pcr Assay ◽  
Specific Primers ◽  
Predictive Values ◽  
Species Specific ◽  
Seminested Pcr

A seminested PCR assay, based on the amplification of the pneumococcal pbp1A gene, was developed for the detection of penicillin resistance in clinical isolates of Streptococcus pneumoniae. The assay was able to differentiate between intermediate (MICs = 0.25 to 0.5 μg/ml) and higher-level (MICs = ≥1 μg/ml) resistance. Two species-specific primers, 1A-1 and 1A-2, which amplified a 1,043-bp region of thepbp1A penicillin-binding region, were used for pneumococcal detection. Two resistance primers, 1A-R1 and 1A-R2, were designed to bind to altered areas of the pbp1A gene which, together with the downstream primer 1A-2, amplify DNA from isolates with penicillin MICs of ≥0.25 and ≥1 μg/ml, respectively. A total of 183 clinical isolates were tested with the pbp1A assay. For 98.3% (180 of 183) of these isolates, the PCR results obtained were in agreement with the MIC data. The positive and negative predictive values of the assay were 100 and 91%, respectively, for detecting strains for which the MICs were ≥0.25 μg/ml and were both 100% for strains for which the MICs were ≥1 μg/ml.

scholarly journals BK Polyomavirus in Renal Transplantation

2014 ◽  
Vol 03 (01) ◽  
Author(s):  
Alice Kennard Prof David W Johnson
Keyword(s):  
Renal Transplantation ◽  
Bk Polyomavirus

URINE CYTOLOGY AND URINE FLOW CYTOMETRY IN RENAL TRANSPLANTATION—A PROSPECTIVE DOUBLE BLIND STUDY1

1995 ◽  
Vol 59 (4) ◽  
pp. 495-500 ◽  
Author(s):  
Isabel Roberti ◽  
Lewis Reisman ◽  
Lewis Burrows ◽  
Kenneth V. Lieberman
Keyword(s):  
Flow Cytometry ◽  
Renal Transplantation ◽  
Urine Flow ◽  
Urine Cytology ◽  
Double Blind ◽  
Urine Flow Cytometry
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