Document Detail


Detection and quantitation of BK virus DNA by real-time polymerase chain reaction in the LT-ag gene in adult renal transplant recipients.
MedLine Citation:
PMID:  16112754     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Determination of polyomavirus BK (BKV) load in urine and plasma has been advocated for monitoring adult renal transplant recipients suffering from BKV-related nephropathy. An "in-house" real-time quantitative PCR assay was developed using the BKV-1/BKV-3 primers set in the large tumor antigen (LT-ag) region to quantitate BK virus loads in plasma and urine in renal transplant patients. This assay was adapted to routine virology laboratory by evaluating two extraction procedures of nucleic acids from urine and plasma, one manual and the other using an automatic extractor, and by evaluating the Light Cycler versus Taqman apparatus. Both the manual and automatic extraction procedures and real-time PCR apparatus were equivalent. The Light Cycler and Taqman instruments allow similarly rapid, accurate, reproducible and specific quantitative detection of the three major BKV subtypes, with a detection limit of 10 BKV DNA copies/ml, and a range from 10(0) to 10(7) copies/ml. Of 855 renal transplant patients, 128 (15%) had BKV DNA in both plasma and urine samples with a mean viral load of 5.1 log/ml in plasma and 6.8 log/ml in urine and in 5 (4%) BKV-associated tubulo-interstitial nephropathy; 332 (39%) BKV DNA was found only in the urine, not in the plasma, without further development of nephropathy and 395 patients had no BKV in plasma and urine. These observations emphasize the usefulness of real-time PCR to assess the BKV load by routine testing, and confirm the need to combine both plasma and urine determinations of the BKV DNA load in order to identify renal transplant patient at high risk for BKV-associated nephropathy.
Authors:
Ali Si-Mohamed; Jérôme Le Goff; Nathalie Désiré; Sarah Maylin; Denis Glotz; Laurent Bélec
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2005-08-19
Journal Detail:
Title:  Journal of virological methods     Volume:  131     ISSN:  0166-0934     ISO Abbreviation:  J. Virol. Methods     Publication Date:  2006 Jan 
Date Detail:
Created Date:  2005-12-12     Completed Date:  2006-04-13     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  8005839     Medline TA:  J Virol Methods     Country:  Netherlands    
Other Details:
Languages:  eng     Pagination:  21-7     Citation Subset:  IM    
Affiliation:
Laboratoire de Microbiologie, Unité de Virologie, Hôpital Européen Georges Pompidou, 20 rue Leblanc, 75908 Paris Cedex 15, France. ali.simohamed@egp.aphp.fr
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MeSH Terms
Descriptor/Qualifier:
Antigens, Viral, Tumor
BK Virus / genetics,  immunology,  isolation & purification*
DNA Primers
DNA, Viral / blood*,  urine*
Humans
Kidney Transplantation / adverse effects
Nephritis, Interstitial / blood,  etiology,  urine
Polymerase Chain Reaction / methods*
Polyomavirus Infections / blood,  etiology,  urine
Tumor Virus Infections / blood,  etiology,  urine
Viral Load
Chemical
Reg. No./Substance:
0/Antigens, Viral, Tumor; 0/DNA Primers; 0/DNA, Viral

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