ABSTRACT
Objectives: Production of extended spectrum beta -lactamases [ESBLs] by clinical isolates of pathogenic E. coli is a very serious therapeutic threat. This study was aimed to investigate the prevalence of ESBLs and associated drug resistance in E. coli isolates from urine and pus, and to report the drift from 2005 to 2009-10
Methodology: Among 173 E. coli isolates, 82 were phenotypically detected as ESBL producers by standard cefotaxime / clavulanic acid and ceftazidime / clavulanic acid disc diffusion tests. Antimicrobial resistance of all ESBL producers was assessed by disc diffusion method. Presence of CTX-M, TEM, SHV and OXA groups was investigated by PCR
Results: The prevalence of ESBL producing E. coli increased significantly from 33.7% in 2005 to 60.0% in 2009-10 [urine: 31.8% to 62.9%; pus: 41.1% to 55.5%]. Resistance to cefotaxime, ceftazidime, ciprofloxacin, gentamicin, nalidixic acid, ticarcillin-clavulanic acid, and trimethoprim-sulfamethoxazole was above 85% in both sets of isolates. Imipenem and Fosfomycin resistance was non-existent in 2005 but ranged from 3-15% in 2009-10. Remarkable increase from 9.5% to 64.7% in urinary tract isolates and from 0 to 55% in pus isolates was observed in colistin sulphate resistance. The dissemination of genes encoding ESBLs was: CTX-M 3.5%; TEM 10.7%; both CTX-M and TEM 3.5% in 2005, and CTX-M 42.5%; TEM 48.1%; both CTX-M and TEM 29.6% in 2009-10
Conclusions: Our results showed very rapid emergence of multidrug resistant ESBL producing E. coli in Pakistan posing a very serious threat in the treatment of nosocomial and community acquired infections
ABSTRACT
Currently fluoroquinolones and cephalosporins are at the forefront of typhoid treatment. The objective of this study was to assess the current drug resistance status of S. Typhi isolates from Faisalabad region by conventional and molecular methods. Drug resistance pattern of 30 clinical isolates [2011] against 8 drugs [nalidixic acid, ciprofloxacin, ofloxacin, gatifloxacin, cephradine, cefixime, ceftriaxone and cefpodoxime] was determined. MICs were noted by E-test. ESBL production was also tested. Relevant drug resistance genes bla[TEM], bla[OXA], gyrA, gyrB, parC, parE, qnrS and qnrA were targeted and QRDR regions of gyrA, gyrB, parC, and parE were sequenced for mutations. Nalidixic acid and ciprofloxacin resistance were seen in 30.0% and 10.0% of isolates respectively. No resistance was detected towards ofloxacin and gatifloxacin. Resistance to cephradine, cefixime, cefpodoxime and ceftriaxone was found in 46.7%, 13.3%, 16.7%, and 3.3% of isolates respectively. In ciprofloxacin resistant isolates a single mutation at codon Ser83 in gyrA gene was detected. A slow increase in ciprofloxacin resistance was indicated. However, newer fluoroquinolones ofloxacin and gatifloxacin are still very effective. Among cephalosporins, ceftriaxone showed promising results but emerging resistance was evident. Fortunately no ESBL producing isolate was detected. No correlation between two groups was detected in emergence of drug resistance
ABSTRACT
Development of a rapid, reliable PCR - based method for molecular identification of Salmonella enterica serovar Paratyphi A directly from blood samples. S. Paratyphi A isolates were used for regular PCR targeting specific region of fliC-a gene. New primers were designed and conditions were optimized for a nested PCR that could be directly applicable on blood samples. The procedure was tested on 70 blood samples from suspected cases of typhoidal infection and comparison made with blood culture. Blood culture was able to diagnose only four patients as infected with S. Paratyphi A. Regular PCR was unable to detect S. Paratyphi A directly from blood where as nested PCR detected S. Paratyphi A in blood of thirteen patients. S. Paratyphi A, which is emerging as a major pathogen can be detected with better sensitivity by nested PCR as compared with blood culture
Subject(s)
Humans , Polymerase Chain Reaction/methods , Diagnosis/instrumentation , Blood , Typhoid Fever/diagnosis , Paratyphoid Fever/diagnosisABSTRACT
Only proven way of transmission of Hepatitis C is through blood. The origin is unknown in nearly half the cases. Pollution is suspected as a cause but it is impossible to prove this relationship directly. We thought that typhoid being a proven pollution related disease, determination of its confection in Hepatitis C patients representing same Socio-economic group would be of interest. A typhoid in Hepatitis C patients can easily be overlooked because symptoms like fever and abdominal discomfort are present in both diseases. Blood samples were collected from three groups of study as mentioned in materials and methods. These samples were processed for 4[th] generation HCV ELISA. PCR for HCV, PCR for typhoid. Blood culture for typhoid and widal test as required [details are given in methodology]. Finally the data thus obtained was analysed and conclusions were drawn. Health Biotechnology Division, National Institute for Biotechnology and Genetic Engineering [NIBGE] and Millat Laboratory Faisalabad. April 2004 to Oct 2004. The parameters included were PCR, blood culture and widal test. There were three groups of study, PCR and ELISA positive patients of Hepatitis C [105] - further subdivided into two groups, with history of exposure to known causes of spread of HCV in last one year [65] and those without such history [40]; clinically diagnosed cases of typhoid [30]; and healthy controls [50]. In the three groups, PCR was positive in 9.5[7.7 and 12.5], 63.3, and 2.0% cases respectively. Figures for blood culture were 4.7[3.1 and 7.5], 33.3, and 0% in the same order, and the respective figures for widal test were 34.2[33.8 and 35.0], 56.6, and 24.0%. The increase in PCR and blood culture positivity in Hepatitis C cases as compared with normal subjects is statistically significant [P< 0.05]. These results clearly suggest that the source of infection for the two diseases is same in many cases, and therefore, provides a strong indication of a relationship between pollution and Hepatitis C