Development and validation of a quantitative real time PCR assay for BK virus.

Several studies have shown that BK viral load in plasma and urine are reliable markers for the detection of BK virus associated nephropathy (BKVAN) in renal transplant patients. Zeptometrix developed a quantitative real time PCR assay based on TaqMan technology for the measurement of BK viral load in plasma and urine.

Considering the high similarity of the nucleotide sequence of the BK virus (BKV) with the JC virus (JCV), we designed this assay to specifically amplify BKV. We determined the viral DNA recovery rate on manual (QIAGEN’s QIAamp DNA Blood Mini Kit) and automated (BioMerieux’s NucliSENS EasyMAG) extraction methods. Cepheid GenExpert was used as control.

The comparison showed a higher viral DNA recovery rate on the automated extraction (61-76% in plasma and 52-65% in urine) as compared to the manual method (49-52% in plasma and 33-56% in urine). Quantitation of the viral load was performed using an external standard curve that was constructed with serial dilution of a plasmid containing the full length of the BKV genome. Commercially available quantitative BKV standards showed good correlation with the plasmid standard. The reproducibility of the assay was determined based on the Ct values of the amplified products as well as in BK copies per milliliter of sample. This assay is linear over a 7 log range (10 to 1 × 10(7) copies per reaction), no cross-reactivity was detected with the closest-related polyomavirus JCV, as well as other viruses that may be found in immunocompromised patients, and human genomic DNA. The limit of detection of the assay is 300 copies per milliliter in both plasma and urine and the limit of quantitation is 1000 copies per milliliter using the Zeptometrix NATtrol BK Virus Linearity Panel (Gentaur Poland).

This real time PCR assay provides a reliable and sensitive method for the quantitation of BKV in plasma and urine samples.

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NATtrol EV Negative Control (6 X 0.5 mL)
MDZ056	Zeptometrix	6 X 0.2 mL	208 EUR
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Description: Please contact Gentaur in order to receive the datasheet of the product.
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NATLAC-6MC	Zeptometrix	6 X 0.5 mL	313 EUR
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NATLAC-6MC-IVD	Zeptometrix	6 X 0.5 mL	333 EUR
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NATRSV-6C-IVD	Zeptometrix	6 X 0.5 mL	242 EUR
NATtrol GBS Positive Control (6 x 0.25 mL)
NATSAG-6L	Zeptometrix	6 x 0.25 mL	272 EUR
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NATSAG-6MC	Zeptometrix	6 X 0.5 mL	313 EUR
NATtrol GBS Positive Control (6 x 0.5 mL)
NATSAG-6MC-IVD	Zeptometrix	6 x 0.5 mL	333 EUR
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Description: Please contact Gentaur in order to receive the datasheet of the product.
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Description: Please contact Gentaur in order to receive the datasheet of the product.
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Description: Please contact Gentaur in order to receive the datasheet of the product.
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NATHIN-STQ	Zeptometrix	1 mL	857 EUR
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NATtrol S. pneumoniae Stock (Quantitative) (1mL)
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NATMRSA-6MC	Zeptometrix	6 X 0.5 mL	315 EUR
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Description: Please contact Gentaur in order to receive the datasheet of the product.
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NATCT/NGNEG-6MC	Zeptometrix	6 X 1 mL	220 EUR
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NATtrol Cytomegalovirus Stock (Quantitative) (1 mL)
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NATADV40-GP	Zeptometrix	0.5 mL	55 EUR
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NATMSSE-6MC	Zeptometrix	6 X 0.5 mL	307 EUR
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Description: Please contact Gentaur in order to receive the datasheet of the product.
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Description: Please contact Gentaur in order to receive the datasheet of the product.
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Description: Please contact Gentaur in order to receive the datasheet of the product.
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Description: Please contact Gentaur in order to receive the datasheet of the product.
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NATtrol CT/NG Negative Control Pack (6 X 1.25 mL)
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Description: Please contact Gentaur in order to receive the datasheet of the product.
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NATtrol Adenovirus Type 03 Stock (Qualitative) (1 mL)
NATADV3-ST	Zeptometrix	1 mL	1281 EUR
NATtrol Adenovirus Type 31 Stock (Qualitative) (1 mL)
NATADV31-ST	Zeptometrix	1 mL	1281 EUR
NATtrol Adenovirus Type 04 Stock (Qualitative) (1 mL)
NATADV4-ST	Zeptometrix	1 mL	1281 EUR
NATtrol Adenovirus Type 40 Stock (Qualitative) (1 mL)
NATADV40-ST	Zeptometrix	1 mL	1281 EUR
NATtrol Adenovirus Type 41 Stock (Qualitative) (1 mL)
NATADV41-ST	Zeptometrix	1 mL	1281 EUR
NATtrol Adenovirus Type 05 Stock (Qualitative) (1 mL)
NATADV5-ST	Zeptometrix	1 mL	1281 EUR
NATtrol Influenza B Virus Stock (Qualitative) (1 mL)
NATFLUB-ST	Zeptometrix	1 mL	1281 EUR
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NATCDI-6L	Zeptometrix	6 x 0.25 mL	272 EUR
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NATADV3-STQ	Zeptometrix	1 mL	857 EUR
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0831042	Zeptometrix	-	693 EUR
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NATCXVA9-6C-IVD	Zeptometrix	6 X 0.5 mL	240 EUR
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NATFLUAB-6C	Zeptometrix	6 x 0.5mL	211 EUR
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NATFLURSV-6C	Zeptometrix	6 x 0.5 mL	221 EUR
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MDZ045	Zeptometrix	6 X 0.5 mL	272.16 EUR
Description: Please contact Gentaur in order to receive the datasheet of the product.
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NATRPC2.1-BIO	Zeptometrix	12 x 0.3mL	529 EUR
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NATEBV-STQ	Zeptometrix	1 mL	857 EUR
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NATFLUAH1(2009)-STQ	Zeptometrix	1 mL	857 EUR
NATtrol Group A Strep Positive Control (6 x 0.25 mL)
NATSPY-6L	Zeptometrix	6 x 0.25 mL	265 EUR
NATtrol Norovirus GI/GII Positive Control (6 x 0.125mL)
NATNOV-6MC	Zeptometrix	6 x 0.125mL	205 EUR
NATtrol Human Hepatitis B Virus (HBV) (400 IU/mL) (1 mL)
NATHBV-0001	Zeptometrix	400 IU/mL 1 mL	45 EUR
NATtrol Human Hepatitis B Virus (HBV) (5,000 IU/mL) (1 mL)
NATHBV-0002	Zeptometrix	5,000 IU/mL 1 mL	160 EUR
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NATHBV-0003	Zeptometrix	50,000 IU/mL 1 mL	508 EUR

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