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R esearch
(Published online:
23-07-2015)
15.
Detection and sequence analysis of accessory
gene regulator genes of Staphylococcus pseudintermedius
isolates -
M. Ananda Chitra, C.
Jayanthy and B. Nagarajan
Veterinary World, 8(7): 902-907
doi:
10.14202/vetworld.2015.902-907
M.
Ananda Chitra:
Department of Veterinary Microbiology, Madras Veterinary College,
Tamil Nadu Veterinary and Animal Sciences
University, Chennai - 600 007, Tamil Nadu, India; m.anandachitra@tanuvas.org.in
C.
Jayanthy:
Department of Veterinary Clinical Medicine, Madras Veterinary
College, Tamil Nadu Veterinary and Animal Sciences University,
Chennai - 600 007, Tamil Nadu, India;
c.jayanthy@tanuvas.org.in
B.
Nagarajan: Department of Veterinary Clinical Medicine, Madras
Veterinary College, Tamil Nadu Veterinary and Animal Sciences
University, Chennai - 600 007, Tamil Nadu, India;
bnvetdr@yahoo.com
Received:
20-02-2015, Revised: 16-06-2015, Accepted: 26-06-2015, Published
online: 23-07-2015
Corresponding author:
M. Ananda Chitra, e-mail: m.anandachitra@tanuvas.org.in
Citation:
Ananda Chitra M,
Jayanthy C, Nagarajan B (2015) Detection and sequence analysis of
accessory gene regulator genes of Staphylococcus
pseudintermedius isolates, Veterinary World 8(7):
902-907.
Abstract
Background: Staphylococcus pseudintermedius (SP) is the
major pathogenic species of dogs involved in a wide variety of
skin and soft tissue infections. The accessory gene regulator (agr)
locus of Staphylococcus aureus has been extensively
studied, and it influences the expression of many virulence genes.
It encodes a two-component signal transduction system that leads
to down-regulation of surface proteins and up-regulation of
secreted proteins during in vitro growth of S. aureus.
The objective of this study was to detect and sequence analyzing
the AgrA, B, and D of SP isolated from canine skin infections.
Materials and Methods: In this study, we have isolated and
identified SP from canine pyoderma and otitis cases by polymerase
chain reaction (PCR) and confirmed by PCR-restriction fragment
length polymorphism. Primers for SP agrA and agrBD
genes were designed using online primer designing software and
BLAST searched for its specificity. Amplification of the agr
genes was carried out for 53 isolates of SP by PCR and
sequencing of agrA, B, and D were carried out for five isolates
and analyzed using DNAstar and Mega5.2 software.
Results: A total of 53 (59%) SP isolates were obtained from 90
samples. 15 isolates (28%) were confirmed to be
methicillinresistant SP (MRSP) with the detection of the mecA
gene. Accessory gene regulator A, B, and D genes were detected
in all the SP isolates. Complete nucleotide sequences of the above
three genes for five isolates were submitted to GenBank, and their
accession numbers are from KJ133557 to KJ133571. AgrA amino acid
sequence analysis showed that it is mainly made of alpha-helices
and is hydrophilic in nature. AgrB is a transmembrane protein, and
AgrD encodes the precursor of the autoinducing peptide (AIP).
Sequencing of the agrD gene revealed that the 5 canine SP
strains tested could be divided into three Agr specificity groups
(RIPTSTGFF, KIPTSTGFF, and RIPISTGFF) based on the putative AIP
produced by each strain. The AIP of SP contains serine and produce
lactone ring structured AIP.
Conclusion: Presence of AgrA, B, and D in all SP isolates
implies the importance of this regulatory system in the virulence
genes expression of the SP bacteria. SP isolates can be typed
based on the AgrD auto-inducible protein sequences as it is being
carried out for typing of S. aureus isolates. However,
further studies are required to elucidate the mechanism of
controlling of virulence genes by agr gene locus in the
pathogenesis of soft tissue infection by SP.
Keywords: accessory gene regulator, dog,
skin infections, Staphylococcus pseudintermedius.
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