Journal of Clinical Virology - Volume 82 - Supplement 1

Abstracts / Journal of Clinical Virology 82S (2016) S1–S142

S103

remain in a latent form in the urogenital epithelial cells. Although

the association of JC virus (JCV) to viral nephropathy (PVAN) is

unclear, the role of BK virus (BKV) is well documented. In RT recip-

ients, reactivation of BKV is an important cause of kidney allograft

loss and the clinical manifestations range fromasymptomatic repli-

cation in the urinary tract to complications such as ureteric stenosis,

transient impairment of renal function, PVAN and irreversible graft

failure. The classical sequence of disease is usually viruria, viremia

and then nephropathy but kidney transplantation guidelines rec-

ommend BKV testing of plasma, not urine, as the most effective

screening approach. InHSCT recipients, hemorrhagic cystitis (HC) is

the most common complication experienced by recipients infected

with BKV. Nevertheless, up to 80% of HSCT recipients are noted to

have BKV viruria but only 10 to 25% of all patients develop clinically

significant HC. The aim of this study was to assess the incidence of

BKV and JCV infections and the diagnostic sequence influence on

HSCT and RT recipients.

Methods:

A long-termprospective study was conducted among

patients who received a HSCT or a RT in our institution, with posi-

tive urine sample for polyomavirus and with a blood sample taken

at the time of the urine sample, received in our laboratory between

November 2013 and September 2015. qPCR was used to detect

BKV and JCV DNA automatically extracted, with LightMix

Kit

on the Roche Diagnostics LightCycler

2.0 Instrument. PVAN was

diagnosed by renal biopsy and demonstration with the immuno-

histochemical staining SV40T antigen.

Results:

43 patients were enrolled in the study, 27male (62.8%),

mean age 50 years old (range 11–69); 15 RT and 28 HSCT. A

total of 147 samples (74 urine, 73 whole blood) were studied. The

median number of samples was 1.7 urine (1–6) and 1.5 blood (1–6)

per patient and the median time since transplantation to poly-

omavirus detection was 210.45 days (12–947). BKV or JCV viruria

was detected in: 6 (40%) and 8 (53.4%) patients respectively after

RT (1 patient presented both BKV and JCV, 6.7%); 17 (60.7%) and

10 (35.7%) patients after HSCT (4 patients, BKV and JCV, 14.3%).

Positive viremia was found in 3 patients (7%), 2 RT (1 BKV and 1

JCV) and 1 HSCT (BKV). Among RT patients, PVAN was diagnosed

in 2 patients, 1 BKV (viruria 95487 copies/ml, viremia nega-

tive) and 1 JCV (viruria >10

copies/ml, viremia 76900 copies/ml),

both without any other coinfection. 10 hematologic patients pre-

sented HC, 6 (>10

copies/ml) BKV viruria, 2 (>10

copies/ml) JCV

viruria, 1 (20000 copies/ml BKV and 37154 JCV viruria) and 1

(350000 copies/ml) BKV viremia and adenovirus viruria. 1 patient

with HC presented BKV viremia (9268 copies/ml).

Conclusions:

Although a small number of PVAN were detected,

the presence of renal failure together with polyomavirus viruria

should be an indication of renal biopsy in order to rule out PVAN,

due to the absence of viremia in some cases. We also detected one

case of JCV PVAN, which should be taken into account. Regarding

the HC, viremia is not an essential factor.

http://dx.doi.org/10.1016/j.jcv.2016.08.205

Abstract no: 77

Presentation at ESCV 2016: Poster 166

Torque teno virus (TTV) in immunosuppressed

host: Performances studies of TTV R-Gene

kit

and donors and recipients kidney samples

genotyping

D. Kulifaj

1 ,

∗

, M. Essig

, F. Meynier

, N. Pichon

E. Munteanu

, R. Moulinas

, M. Joannes

D. Heckel

, J. Combrissson

, C. Barranger

S. Alain

bioMerieux, 138 rue Louis Pasteur, Parc

Technologique Delta Sud, 09340 Verniolle,

France-INSERM UMR1092, CNR CMV, CHU LIMOGES,

France

Service de Néphrologie, CHU de Limoges

bioMerieux, Centre Christophe Mérieux, 5 rue des

berges, 38024 Grenoble Cedex 01, France

Reanimation, Chu de Limoges

INSERM UMR1092, CNR CMV, CHU LIMOGES,

France - Service de Néphrologie, CHU de Limoges

Genomic platform GenoLim, Limoges University

bioMerieux, 138 rue Louis Pasteur, Parc

Technologique Delta Sud, 09340 Verniolle, France

INSERM UMR1092, CNR CMV, CHU LIMOGES,

France - Genomic platform GenoLim, Limoges

University

Background:

TTV are highly prevalent in the general popula-

tion and infections are characterized by lifelong viremia. These

viruses are so far not associated with clinical disease, but associ-

ation of viral load variation and immunosuppression has recently

been reported. Though, clarifying the role of TTV as a surrogate

marker and evaluating the interest of viral load followup tomonitor

patient immunity requires standardized PCR systems that accu-

rately detect, and quantify, all the human genotypes.

We thus developed the TTV R-gene

kit (ARGENE

range,

bioMérieux, France, available soon) for the detection and quantifi-

cation of TTV in whole blood and studied its performances as an

immunomodulation marker. TTV is also a virus with a high genetic

variability, and the TTV population of genotypes, their transmission

and their evolution in our patientsmay reflect the clinical condition

of immunosuppressed patients.

Methods:

Analytical studies were performed on whole blood

dilutions of quantified TTV samples. The samples were extracted

by NucliSENS

easyMAG

(bioMerieux) and amplified by TTV R-

gene

kit with 10 L of eluate and 15 L of premix ready-to-use.

911 samples (from 42 kidney transplant recipients follow-up),

32 samples from donors were analyzed with the TTV R-gene

. 66

samples of kidneys donors and recipients (paired) were prepro-

cessed and extracted before RCA amplification and purification. The

libraries were prepared with Ion express library kit and sequenced

by Ion Proton high throughput sequencing technology (ion Torrent,

Life technologies).

Results:

LoD:

The limit of detection in whole blood determined by Probit

analysis is 148 cp/mL of sample on whole blood.

Linearity:

Linearity of the quantification results is demonstrated

over the range 2.1

10E + 3 to 4.1

10E + 10 cp/mL.

Specificity:

All TTV genotypes were correctly detected and quan-

tified in whole blood samples with TTV R-gene

. The following

viruses were not detected with TTV R-gene

kit: animal TTV, CMV,

EBV, BK, Adenovirus, HSV1/2, VZV, HHV6/7/8 and B19.

Precision:

The coefficient of total variability ranged between 2.6%

(upper limit) and 11.4% (lower limit).