Abstract: Legionella pneumophila is involved in more than 95%
cases of severe atypical pneumonia. Infection is mainly by
inhalation the indoor aerosols through the water-coolant systems.
Because some Legionella strains may be viable but not culturable,
therefore, Taq polymerase, DNA amplification and semi-nested-PCR
were carried out to detect Legionella-specific 16S-rDNA sequence.
For this purpose, 1.5 litter of water samples from 77 water-coolant
system were collected from four different hospitals, two nursing
homes and one student hostel in Kerman city of Iran, each in a brand
new plastic bottle during summer season of 2006 (from April to
August). The samples were filtered in the sterile condition through
the Millipore Membrane Filter. DNA was extracted from membrane
and used for PCR to detect Legionella spp. The PCR product was
then subjected to semi-nested PCR for detection of L. pneumophila.
Out of 77 water samples that were tested by PCR, 30 (39%) were
positive for most species of Legionella. However, L. pneumophila
was detected from 14 (18.2%) water samples by semi-nested PCR.
From the above results it can be concluded that water coolant
systems of different hospitals and nursing homes in Kerman city of
Iran are highly contaminated with L. pneumophila spp. and pose
serious concern. So, we recommend avoiding such type of coolant
system in the hospitals and nursing homes.
Abstract: Fourty one strains of ESBL producing P.aeruginosa
which were previously isolated from burn patients in Kerman
University general hospital, Iran were subjected to PCR, RFLP and
sequencing in order to determine the type of extended spectrum β-
lactamases (ESBL), the restriction digestion pattern and possibility of
mutation among detected genes. DNA extraction was carried out by
phenol chloroform method. PCR for detection of bla genes was
performed using specific primer for each gene. Restriction Fragment
Length Polymorphism (RFLP) for ESBL genes was carried out using
EcoRI, NheI, PVUII, EcoRV, DdeI, and PstI restriction enzymes. The
PCR products were subjected to direct sequencing of both the strands
for identification of the ESBL genes.The blaCTX-M, blaVEB-1, blaPER-1,
blaGES-1, blaOXA-1, blaOXA-4 and blaOXA-10 genes were detected in the
(n=1) 2.43%, (n=41)100%, (n=28) 68.3%, (n=10) 24.4%, (n=29)
70.7%, (n=7)17.1% and (n=38) 92.7% of the ESBL producing isolates
respectively. The RFLP analysis showed that each ESBL gene has
identical pattern of digestion among the isolated strains. Sequencing
of the ESBL genes confirmed the genuinety of PCR products and
revealed no mutation in the restriction sites of the above genes. From
results of the present investigation it can be concluded that blaVEB-1
and blaCTX-M were the most and the least frequently isolated ESBL
genes among the P.aeruginosa strains isolated from burn patients. The
RFLP and sequencing analysis revealed that same clone of the bla
genes were indeed existed among the antibiotic resistant strains.