Abstract: Polymerase chain reaction (PCR) assays targeting genomic DNA segments have been established for the detection of Helicobacter pylori in clinical specimens. However, the data on comparative evaluations of various targets in detection of H. pylori are limited. Furthermore, the frequencies of vacA (s1 and s2) and cagA genotypes, which are suggested to be involved in the pathogenesis of H. pylori in other parts of the world, are not well studied in Kuwait. The aim of this study was to evaluate PCR assays for the detection and genotyping of H. pylori by targeting the amplification of DNA targets from four genomic segments. The genomic DNA were isolated from 72 clinical isolates of H. pylori and tested in PCR with four pairs of oligonucleotides primers, i.e. ECH-U/ECH-L, ET-5U/ET-5L, CagAF/CagAR and Vac1F/Vac1XR, which were expected to amplify targets of various sizes (471 bp, 230 bp, 183 bp and 176/203 bp, respectively) from the genomic DNA of H. pylori. The PCR-amplified DNA were analyzed by agarose gel electrophoresis. PCR products of expected size were obtained with all primer pairs by using genomic DNA isolated from H. pylori. DNA dilution experiments showed that the most sensitive PCR target was 471 bp DNA amplified by the primers ECH-U/ECH-L, followed by the targets of Vac1F/Vac1XR (176 bp/203 DNA), CagAF/CagAR (183 bp DNA) and ET-5U/ET-5L (230 bp DNA). However, when tested with undiluted genomic DNA isolated from single colonies of all isolates, the Vac1F/Vac1XR target provided the maximum positive results (71/72 (99% positives)), followed by ECH-U/ECH-L (69/72 (93% positives)), ET-5U/ET-5L (51/72 (71% positives)) and CagAF/CagAR (26/72 (46% positives)). The results of genotyping experiments showed that vacA s1 (46% positive) and vacA s2 (54% positive) genotypes were almost equally associated with VaCA+/CagA- isolates (P > 0.05), but with VacA+/CagA+ isolates, S1 genotype (92% positive) was more frequently detected than S2 genotype (8% positive) (P< 0.0001). In conclusion, among the primer pairs tested, Vac1F/Vac1XR provided the best results for detection of H. pylori. The genotyping experiments showed that vacA s1 and vacA s2 genotypes were almost equally associated with vaCA+/cagA- isolates, but vacA s1 genotype had a significantly increased association with vacA+/cagA+ isolates.
Abstract: New diagnostic tools are urgently needed to interrupt the transmission of tuberculosis and multidrug-resistant tuberculosis. The microscopic-observation drug-susceptibility (MODS) assay is a rapid, accurate and simple liquid culture method to detect multidrug-resistant tuberculosis (MDR-TB). MODS were evaluated to determine a lower and same concentration of isoniazid and rifampin for detection of MDR-TB. Direct drug-susceptibility testing was performed with the use of the MODS assay. Drug-sensitive control strains were tested daily. The drug concentrations that used for both isoniazid and rifampin were at the same concentration: 0.16, 0.08 and 0.04μg per milliliter. We tested 56 M. tuberculosis clinical isolates and the control strains M. tuberculosis H37RV. All concentration showed same result. Of 53 M. tuberculosis clinical isolates, 14 were MDR-TB, 38 were susceptible with isoniazid and rifampin, 1 was resistant with isoniazid only. Drug-susceptibility testing was performed with the use of the proportion method using Mycobacteria Growth Indicator Tube (MGIT) system as reference. The result of MODS assay using lower concentration was significance (P
Abstract: A total of twenty tensile biopsies were collected from
children undergoing tonsillectomy from teaching hospital ENT
department and Kurdistan private hospital in sulaimani city. All
biopsies were homogenized and cultured; the obtained bacterial
isolates were purified and identified by biochemical tests and VITEK
2 compact system. Among the twenty studied samples, only one
Pseudomonas putida with probability of 99% was isolated.
Antimicrobial susceptibility was carried out by disk diffusion
method, Pseudomonas putida showed resistance to all antibiotics
used except vancomycin. The isolate further subjected to PCR and
DNA sequence analysis of blaVIM gene using different set of primers
for different regions of VIM gene. The results were found to be PCR
positive for the blaVIM gene. To determine the sequence of blaVIM
gene, DNA sequencing performed. Sequence alignment of blaVIM
gene with previously recorded blaVIM gene in NCBI- database showed
that P. putida isolate have different blaVIM gene.
Abstract: Antimicrobial resistant is becoming a major factor in
virtually all hospital acquired infection may soon untreatable is a
serious public health problem. These concerns have led to major
research effort to discover alternative strategies for the treatment of
bacterial infection. Nanobiotehnology is an upcoming and fast
developing field with potential application for human welfare. An
important area of nanotechnology for development of reliable and
environmental friendly process for synthesis of nanoscale particles
through biological systems In the present studies are reported on the
use of fungal strain Aspergillus species for the extracellular synthesis
of bionanoparticles from 1 mM silver nitrate (AgNO3) solution. The
report would be focused on the synthesis of metallic bionanoparticles
of silver using a reduction of aqueous Ag+ ion with the
culture supernatants of Microorganisms. The bio-reduction of the
Ag+ ions in the solution would be monitored in the aqueous
component and the spectrum of the solution would measure through
UV-visible spectrophotometer The bionanoscale particles were
further characterized by Atomic Force Microscopy (AFM), Fourier
Transform Infrared Spectroscopy (FTIR) and Thin layer
chromatography. The synthesized bionanoscale particle showed a
maximum absorption at 385 nm in the visible region. Atomic Force
Microscopy investigation of silver bionanoparticles identified that
they ranged in the size of 250 nm - 680 nm; the work analyzed the
antimicrobial efficacy of the silver bionanoparticles against various
multi drug resistant clinical isolates. The present Study would be
emphasizing on the applicability to synthesize the metallic
nanostructures and to understand the biochemical and molecular
mechanism of nanoparticles formation by the cell filtrate in order to
achieve better control over size and polydispersity of the
nanoparticles. This would help to develop nanomedicine against
various multi drug resistant human pathogens.
Abstract: Fungal infections are becoming more common and the
range of susceptible individuals has expanded. While Candida
albicans remains the most common infective species, other Candida
spp. are becoming increasingly significant. In a range of large-scale
studies of candidaemia between 1999 and 2006, about 52% of 9717
cases involved C. albicans, about 30% involved either C. glabrata or
C. parapsilosis and less than 15% involved C. tropicalis, C. krusei or
C. guilliermondii. However, the probability of mortality within 30
days of infection with a particular species was at least 40% for C.
tropicalis, C. albicans, C. glabrata and C. krusei and only 22% for
C. parapsilopsis. Clinical isolates of Candida spp. grew at rates
ranging from 1.65 h-1 to 4.9 h-1. Three species (C. krusei, C. albicans
and C. glabrata) had relatively high growth rates (μm > 4 h-1), C.
tropicalis and C. dubliniensis grew moderately quickly (Ôëê 3 h-1) and
C. parapsilosis and C. guilliermondii grew slowly (< 2 h-1). Based
on these data, the log of the odds of mortality within 30 days of
diagnosis was linearly related to μm. From this the underlying
probability of mortality is 0.13 (95% CI: 0.10-0.17) and it increases
by about 0.09 ± 0.02 for each unit increase in μm. Given that the
overall crude mortality is about 0.36, the growth of Candida spp.
approximately doubles the rate, consistent with the results of larger
case-matched studies of candidaemia.
Abstract: Studies were carried out to determine the in vitro
susceptibility of the typhoid pathogens to combined action of Euphorbia hirta, Euphorbia heterophylla and Phyllanthus niruri. Clinical isolates of the typhoid bacilli were subjected to susceptibility testing using agar diffusion technique and the minimum inhibitory
concentration (MIC) determined with tube dilution technique. These
isolates, when challenged with doses of the extracts from the three
medicinal plants showed zones of inhibition as wide as 26±0.2mm, 22±0.1mm and 18±0.0mm respectively. The minimum inhibitory concentration (MIC) revealed organisms inhibited at varying
concentrations of extracts: E. hirta (S. typhi 0.250mg/ml, S. paratyphi A 0.125mg/ml, S. paratyphi B 0.185mg/ml and S. paratyphi C 0.225mg/ml), E. heterophylla (S. typhi 0.280mg/ml, S. paratyphi A
0.150mg/ml, S. paratyphi B 0.200mg/ml and S. paratyphi C 0.250mg/ml) and P. niruri (S. typhi 0.150mg/ml, S. paratyphi A 0.100mg/ml, S. paratyphi B 0.115mg/ml and S. paratyphi C 0.125mg/ml). The results of the synergy between the three plants in
the ration of 1:1:1 showed very low MICs for the test pathogens as follows S. typhi 0.025mg/ml, S. paratyphi A 0.080mg/ml, S. paratyphi B 0.015mg/ml and S. paratyphi C 0.10mg/ml with the
diameter zone of inhibition (DZI) ranging from 35±0.2mm,
28±0.4mm, 20±0.1mm and 32±0.3mm respectively. The secondary
metabolites were identified using simple methods and HPLC. Organic components such as anthroquinones, different alkaloids,
tannins, 6-ethoxy-1,2,3,4-tetrahydro-2,2,4-trimethyl and steroids were identified. The prevalence of Salmonellae, a deadly infectious disease, is still very high in parts of Nigeria. The synergistic action of these three plants is very high. It is concluded that pharmaceutical companies should take advantage of these findings to develop new
anti-typhoid drugs from these plants.