RAPID DETECTION OF HUMAN TORQUE TENO VIRUSES
USING HIGH-RESOLUTION MELTING ANALYSIS
Spandole S1*, Cimponeriu D1, Toma M1, Radu I1, Ion DA2
*Corresponding Author: Ms. Sonia Spandole (Ph.D. Student), Department of Genetics, University of Bucharest,
Intrarea Portocalelor Street, No 1-3, 060101, Bucharest, Romania; Tel.: 004-0764-824-281, Fax: 004-0213-181-
565; E-mail: sonia.spandole@gmail.com
page: 55
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Abstract
Torque teno viruses (TTVs) are recently discovered
DNA viruses, with heterogeneous genomes,
highly prevalent in populations worldwide. The species
that infect humans are Torque teno virus (TTV),
Torque teno midi virus (TTMDV) and Torque teno
mini virus (TTMV). High-resolution melting analysis
(HRMA) is a sensitive and effective method for genotyping
and mutation scanning. Up to now, HRMA
has not been utilized for detection of TTVs.
The aim of this study was to asses if HRMA
is suitable for detecting TTVs variants. DNA was
extracted from the blood and saliva of 13 healthy
subjects for method optimization. Additionally, saliva
samples from 100 healthy individuals were collected
for estimating the TTVs’ prevalence. Viral DNA was
amplified by heminested polymerase chain reaction
(PCR). Second round amplicons were used for the
HRMA. The samples were analyzed using two fluorescent
dyes, SYBR® Green I and EvaGreen®.
The prevalence values for TTV, TTMDV and
TTMV were 71.0, 31.0 and 54.0%, respectively. The
three major melting curve patterns corresponding to
TTV, TTMDV and TTMV on HRMA can be easily
distinguished regardless of kit used. Our results
showed that HRMA is a rapid and efficient method
of detecting human TTVs.
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