Surveillance of HIV infection in blood donors in Pakistan: a systematic review

Objective: To determine the prevalence of human immunodeficiency virus [HIV] in blood donors in a tertiary care hospital blood bank and then compared with prevalence in other reported studies in Pakistan between 1988 and 2016. Study Design: Prospective single centre study with systematic review. Place and Duration of Study: Department of Blood Bank, Shaheed Zulfiqar Ali Bhutto Medical University, PIMS Hospital Islamabad, from Jan 2015 to Dec 2016

Material and Methods: Blood donors were screened for the prevalence of HIV, from Jan 2015 to Dec 2016. Metaanalyses were gathered from the reported HIV incidence in blood donors from across Pakistan during 1988-2016 by searching through Google, Pub Med, and Pak Medi Net [for Pakistani non-indexed journals]

Results: A total of 54,877 blood donors were screened for HIV by Chemiluminescent immunoassay [CLIA], of which 75 [0.13%] male donors were found reactive for HIV. The mean prevalence of HIV from 2006-2016 was 0.06% while 0.13% in 47 studies conducted during the period 1988-2016

Conclusion: The prevalence of HIV in apparently healthy blood donors is steadily increasing in Pakistan. Concerted efforts need to be made to ensure that blood is adequately screened for HIV in every blood bank in the country

In vitro efficacy of meropenem, colistin and tigecycline against the extended spectrum beta-lactamase producing gram negative bacilli

To compare the in vitro efficacy of meropenem, colistin and tigecycline against extended spectrum Beta-lactamase producing Gram negative bacilli by minimal inhibitory concentration. Cross-sectional descriptive study. Department of Microbiology, Army Medical College, National University of Sciences and Technology, Rawalpindi, from June to December 2010. Routine clinical specimens were subjected to standard microbiological procedures and the isolates were identified to species level. Extended spectrum beta-lactamase producing Gram negative bacilli were detected by Jarlier disc synergy method and confirmed by ceftazidime and ceftazidime-clavulanate Etest. Minimum Inhibitory Concentration [MIC[90]] of meropenem, colistin and tigecycline was determined by Etest [AB BIOMERIUX] and the results were interpreted according to the manufacturer's instructions and Clinical and Laboratory Standards Institute guidelines and Food and Drug Authority recommendations. Results were analyzed by using Statistical Package for the Social Sciences version 20. A total of 52 non-duplicate extended spectrum Beta-lactamase-producing Gram negative bacilli were included in the study. The MIC[90] of tigecycline [0.75 microg/ml] was lowest as compared to the meropenem [2 microg/ml] and colistin [3 microg/ml]. Tigecycline is superior in efficacy against the extended spectrum Beta-lactamase producing Gram negative bacilli as compared to colistin and meropenem

Carbapenem resistant acinetobacter-A major pathogen in a Tertiary Care Hospital

Objective: To determine the frequency of carbapenem resistant Acinetobacter in the Military Hospital Rawalpindi, Pakistan

Study Design: Descriptive study

Place and Duration of Study: Department of Microbiology, Army Medical College, from Oct, 2012 to Feb, 2013

Material and Methods: Clinical specimens like naso-bronchial lavage, blood, pus, sputum and catheter tips were inoculated on blood agar and Mac Conkey agar while the urine samples were inoculated on Cystine Lactose Electrolyte Deficient [CLED] agar. Acinetobacter spp. isolated, were later subjected to antimicrobial susceptibility testing using the modified Kirby-Bauer disc diffusion method on Mueller Hinton agar as per Clinical Laboratory and Standards Institute [CLSI] guidelines

Results: Out of a total of 85 Acinetobacter spp. 62 isolates were found to be carbapenem resistant. They were also found to be 100% resistant to ciprofloxacin and ceftriaxone thus becoming multidrug resistant followed by tazobactam piperacillin [98%] and trimethoprim sulphamethoxazole [92%]. Minimum resistance was seen against tigecycline being 21%

Conclusion: It is concluded from our study that there is a high frequency [72.94%] of resistance to carbapenems in Acinetobacter spp. in our setup which is associated with increased morbidity and mortality due to limited treatment options

Coagulase negative staphylococci - a fast emerging threat

To determine the frequency of isolation of coagulase-negative staphylococci and their resistance to methicillin over a period of time. The descriptive cross-sectional study was carried out at Army Medical College, Rawalpindi, from June 2009 to May 2012, and comprised clinical samples mostly from patients admitted to the intensive care unit. They were inoculated onto appropriate culture media depending upon the specimen. After 24-hour incubation at 35°C, coagulase-negative staphylococci were identified on the basis of colony morphology, gram staining, a positive catalase and a negative tube coagulase test. Methicillin resistance among the isolated staphylococci was determined using a 30microg Cefoxitin disc as per the Clinical and Laboratory Standards Institute protocol. Number of coagulase-negative staphylococci for each year and their methicillin resistance rates were calculated. A comparison was made with methicillin resistant staphylococcus aureus] isolated during the same period. Of the total 1331 specimens studies over three years, 581[43.65%] were coagulase-negative staphylococci. The rate of coagulase-negative staphylococci and methicillin resistance was higher each year; 110[26.6%] in May 2009-Jun 2010, 134[36.5%] in 2011, and 337[61%] in 2012. Methicillin resistance rates also increased from 25[22.7%] to 46[34.3%] and then to 201[59.6%] in 2012.Maximum isolated specimens came from blood 311[53.5%], followed by pus/swabs 204[35.1%]. The frequency of isolation of coagulase-negative staphylococci and its methicillin resistance among hospitalised patients is on the rise

Frequency and antibiogram of vancomycin resistant enterococcus in a tertiary care hospital

To determine the frequency of Vancomycin Resistant Enterococcus [VRE] in a tertiary care hospital of Rawalpindi, Pakistan. Observational, cross-sectional study. Department of Microbiology, Army Medical College, Rawalpindi, from May 2011 to May 2012. Vancomycin resistant Enterococcus isolated from the clinical specimens including blood, pus, double lumen tip, ascitic fluid, tracheal aspirate, non-directed bronchial lavage [NBL], cerebrospinal fluid [CSF], high vaginal swab [HVS] and catheter tips were cultured on blood agar and MacConkey agar, while the urine samples were grown on cystine lactose electrolyte deficient agar. Later the antimicrobial susceptibility testing of the isolates was carried out using the modified Kirby-Bauer disc diffusion method on Mueller Hinton agar. A total of 190 enterococci were isolated. Of these, 22 [11.57%] were found to be resistant to vancomycin. The antimicrobial sensitivity pattern revealed maximum resistance against ampicillin [86.36%] followed by erythromycin [81.81%] and gentamicin [68.18%] while all the isolates were 100% susceptible to chloramphenicol and linezolid. The frequency of VRE was 11.57% with the highest susceptibility to linezolid and chloramphenicol

In vitro efficacy of Colistin against multi-drug resistant Pseudomonas aeruginosa by minimum inhibitory concentration

Multi-drug resistant bacteria are an important cause of mortality and morbidity. In the management of various infections, timely detection and appropriate treatment, in accordance with the culture and sensitivity reports can help improve the treatment outcome. Colistin is a bactericidal antibiotic which is emerging as a reliable solution for treating infections with multi-drug resistant Gram negative bacilli. The aim of this study was to find out the in-vitro efficacy of colistin against multidrug resistant Pseudomonas aeruginosa isolates by minimum inhibitory concentration. This cross sectional, descriptive study was conducted in the Department of Microbiology, Army Medical College, National University of Sciences and Technology, Islamabad from February 2010 to January 2011. Antimicrobial sensitivity testing was done on Pseudomonas aeruginosa isolated from routine clinical specimens received and the strains which appeared resistant to at least one antimicrobial agent in three or more anti-pseudomonal antimicrobial categories were subjected to the Colistin Etest. The MIC endpoint of colistin was read, as per manufacturer's instructions [AB Biodisk, Solna, Sweden]. The isolates showing MIC of 2micro.g/ml or less were considered sensitive, those with 4-6 micro.g/ml as intermediate and >/= 8 micro.gg/ml as resistant. MIC[50] and MIC[90] of colistin against MDR Pseudomonas aeruginosa was determined. A total of 52 MDR Pseudomonas aeruginosa strains were isolated during the period of the study. The highest percentage was isolated from urine [36%] followed by respiratory tract infections [18%] and pus specimens [20%]. The highest percentage of these isolates was found to be susceptible to colistin followed by piperacillin-tazobactam and cefoperazone-sulbactam. A total of 36 [69%] isolates were sensitive, 10 [20%] were intermediate and 6 [11%] were resistant to colistin by Kirby Bauer disc diffusion method. MIC[50] was found to be l.0 micro.gg/ml while MIC[90] was 3.0 micro.gg/ml. Colistin is a reliable solution in cases of infections with MDR, XDR or PDR Pseudomonas aeruginosa

Evaluation of different phenotypic methods for detection of amp C beta-lactamase producing bacteria in clinical isolates

To compare the sensitivity and specificity of different phenotypic methods for detection of Amp C beta-lactamase producing bacteria. Analytical study. Department of Microbiology, Army Medical College / National University of Sciences and Technology [NUST], Islamabad, Pakistan, from June 2010 to December 2010. A total of 150 clinical isolates were screened for presence of Amp C beta-lactamase by using the cefoxitin disc. The confirmatory methods evaluated were inhibitor based assay [boronic acid], Amp C disc test and Amp C Etest. Three dimensional enzyme extract assay was used as the reference method for determining the sensitivity and specificity. Among the total isolates tested, 62.8% bacteria showed the presence of Amp C beta-lactamase by standard three dimensional enzyme extract assay. Among the three methods compared, boronic acid disk test found out to be highly sensitive [88%] and specific [92%] for the detection of Amp C beta-lactamase producing bacteria. Detection of Amp C production is crucial in order to establish the antibiotic therapy and to attain the favourable clinical outcomes. Implementation of simple tests like boronic acid disk tests in the laboratories will help to alleviate the spread of Amp C beta-lactamase harboring organisms

Evaluation of combined disc method for the detection of metallo-β-lactamase producing Gram negative bacilli

Aims: Infections due to metallo-β-lactamase (MBL) producing Gram negative rods are a cause of high mortality and morbidity. Early detection by an economical and accurate method may improve patient outcome. This study was aimed to evaluate the diagnostic accuracy of combined disc method for MBL detection by comparing it with MBL-Etest. Methodology and Results: This cross-sectional, validation study was carried out in the Department of Microbiology, Army Medical College, National University of Sciences and Technology, Rawalpindi, over a period of six months. A total of 52 non-duplicate Gram-negative rods isolated from the routine clinical specimens and found resistant to meropenem/imipenem on Kirby Bauer Disc Diffusion method were subjected to two tests for metallo-β-lactamase detection. One was combined Disc test using imipenem with Ethylene Diamine Tetraacetic Acid (EDTA), where a strain showing an increase in zone of inhibition of combined disc of ≥ 7 mm as compared to imipenem alone, was considered as MBL producer and the other one was MBL-Etest for which results were interpreted as per manufacturer’s guidelines. Combined disc method for MBL detection was found to have a sensitivity, specificity, positive predictive value, negative predictive value and accuracy of 97.5%, 100%, 100%, 92% and 98%. Conclusion, Significance and Impact of study: Combined disc method is an economical and reliable method for metallo-β-lactamase detection which can be used routinely in any laboratory.

Detection and antibiotic susceptibility pattern of biofilm producing Gram positive and Gram negative bacteria isolated from a tertiary care hospital of Pakistan

Microorganisms adhere to non-living material or living tissue, and form biofilms made up of extracellular polymers/slime. Biofilm-associated microorganisms behave differently from free-floating bacteria with respect to growth rates and ability to resist antimicrobial treatments and therefore pose a public health problem. The objective of this study is to detect the prevalence of biofilm producers among Gram positive and Gram negative bacteria isolated from clinical specimens, and to study their antimicrobial susceptibility pattern. The study was carried out from October 2009 to March 2010, at the Department of Microbiology, Army Medical College/ National University of Sciences and Technology (NUST), Rawalpindi, Pakistan. Clinical specimens were received from various wards of a tertiary care hospital. These were dealt by standard microbiological procedures. Gram positive and Gram negative bacteria isolated were subjected to biofilm detection by congo red agar method (CRA). Antimicrobial susceptibility testing of those isolates, which showed positive results (slime production), was done according to the Kirby-Bauer disc diffusion technique. A total of 150 isolates were tested for the production of biofilm/slime. Among them, 81 isolates showed positive results. From these 81, 51 were Gram positive and 30 were Gram negative. All the 81(54%) slime producers showed reduced susceptibility to majority of antibiotics. Bacterial biofilms are an important virulence factor associated with chronic nosocomial infection. Detection of biofilm forming organisms can help in appropriate antibiotic choice.

Frequency and antibiogram of multi-drug resistant pseudomonas aeruginosa

To find out the frequency and susceptibility pattern of multi-drug resistant [MDR] Pseudomonas aeruginosa in clinical specimens. Cross-sectional observational study. Department of Microbiology, Army Medical College, National University of Sciences and Technology [NUST], Rawalpindi, from January to September 2010. Routine clinical specimens were subjected to standard microbiological procedures and the isolates were identified to the species level. The antibiotics susceptibility was determined by Kirby Bauer Disc diffusion method and the results were interpreted according to the Clinical and Laboratory Standards Institute [CLSI] guidelines. The frequency of MDR Pseudomonas aeruginosa among all the Pseudomonas aeruginosa strains isolated was found to be 22.7%. These isolates were most sensitive to Colistin followed by Piperacillin-Tazobactam and Cefoperazone-Sulbactum. Increasing fequency of infections due to MDR Pseudomonas aeruginosa is an emerging threat in our set up which can be prevented by prescribing antibiotics judiciously and by adopting proper disinfection measures

Antenatal screening for hepatitis B and C virus infection in pregnant women in a tertiary care hospital of Rawalpindi

To determine the frequency of Hepatitis B and C virus infection in pregnant women and to assess whether it is more common in multigravida as compared to primigravida. A Descriptive Cross Sectional study carried out in the out patient department of Gynaecology and Obstetrics department of Military Hospital, Rawalpindi from August to October 2010. An open and close-ended questionnaire was self designed for this purpose. Data relating to medical, surgical, gynaecological and obstetric history, with particular references to hospital admissions and intravenous interventions was collected through convenience sampling. One hundred and forty pregnant women were questioned. The age of patients varied from 21 to 45 years [mean 28.66 years]. Out of 140 females, 44 [31.4%] females were primigravidas while 96 [68.6%] were multigravidas. Eight [5.7%] females were found positive for HBsAg and 18 [12.8%] were found positive for HCV antibodies by third generation ELISA. It was observed that 0% primigravida and 8.3% cases of multigravida were HBsAg positive cases [p = 0.049]. Frequency of HCV was 4.5% in primigravidas and 16.7% in multigravidas [p = 0.047].Various risk factors like, history of blood transfusion 27.1%, and IV injection 91.4%, were identified. The hepatitis B vaccination rate was only 24.3%. The frequency of Hepatitis B and C infection was 5.7% and 12.8% respectively in pregnant females as compared to the general population. The multigravidas were at a higher risk of HBV and HCV exposure than primigravida

Frequency and antibiotic susceptibility pattern of Amp C beta-lactamase producing bacteria isolated from a tertiary care hospital of Rawalpindi, Pakistan

Amp C beta-lactamases are cephalosporinases which hydrolyze cephamycins and are poorly inhibited by clavulanic acid. Amp C beta-lactamases confer resistance to a wide variety of antibiotics and pose both diagnostic and therapeutic challenges. The objective was to detect the frequency and antibiotic susceptibility pattern of Amp C beta-lactamase producing bacteria isolated from a tertiary care hospital of Pakistan. Organisms were isolated from various clinical specimens. First, the screening of the isolates was done by using cefoxitin disc. Screen-positive organisms were subjected to three dimensional extract test for detection of Amp C beta-lactamases. From a total of 100 organism tested, 64 organisms were positive on cefoxitin screen test. Out of these 64, 40 [62.5%] showed the presence of Amp C beta-lactamase. [E.coli n=18, K.pneumoniae n=14, K.oxytoca n=1, Enterobacter species n=5, Citrobacter freundii n=2] by three dimensional extract test. The antibiotics found out to show good activity against these resistant bacteria include meropenem and tigecycline. This is the first study to determine the frequency of Amp C beta lactamases from Pakistan. This study shows a high frequency of Amp C beta-lactamase producing isolates from a hospital, which may lead to serious therapeutic problems

Antimicrobial activity of tigecycline against methicillin resistant staphylococcus aureus in a tertiary care setting

To determine the in vitro efficacy of tigecycline against methicillin resistant Staphylococcus aureus [MRSA]. Department of Microbiology Army Medical College and Armed Forces Institute of Pathology Rawalpindi, from Feb 2008 to Jan 2009. One hundred clinical isolates of MRSA were taken, Detection of MRSA was done using 30 microg disc of cefoxitin as recommended by Clinical laboratory Standard Institute [CLSI]. Susceptibility of the isolates to tigecycline was done by employing modified Kirby Bauer disk diffusion technique, according to the guidelines provided by the Food and Durg Administration [FDA]. Minimum inhibitory concentrations [MICs] of the isolates were determined by using E-strips [bioMerieux] of tigecycline. Results were interpreted according to FDA recommendations. All MRSA isolates were susceptible to tigecycline by disc diffusion method. The MICs of tigecycline revealed that all MRSA isolates were in sensitive range. In an era of rapidly growing antibiotic resistance, tigecycline has been found to have very good in vitro efficacy against MRSA isolates

Relative changes in hepatitis-C virus RNA titre and serum ALT levels before, during and after treatment with interferon and ribavirin

Hepatitis C is commonly treated with Interferon and Ribavirin combination therapy. The aim of this study was to find the relative change in the Hepatitis C virus [HCV] RNA titre and serum alamine aminotransferase [ALT] levels before, during and after completion of treatment with Interferon [IFN] and Ribavirin therapy in HCV positive patients. This cross- sectional descriptive study was conducted from May 2010 to August 2010. Seventy hepatitis C patients reporting to the Laboratory of Army Medical College, Rawalpindi, were included in the study. Blood samples were first subjected to centrifugation and stored at -20[degree sign] C till tested. HCV RNA titre was determined by Real Time PCR while ALT estimation on serum samples was done on Selectra-E by kinetic method. The age, gender and treatment status of the patients was also recorded. In a total of 70 patients, 30% were females while 70% were males. The mean age of patients was 38 +/- 11 yrs. On applying, analysis of variance [ANOVA], we found that the change in serum ALT level was significant [F = 6.044 and p = 0.004] while it was not significant in case of HCV RNA [F = 1.93 and p = 0.153] treatment with Inj. IFN and Ribavirin. There is a definite decreasing trend in the serum ALT levels while no definite relationship of the HCV RNA level to the treatment with combination therapy with IFN and Ribavirin

Awareness of novel h1n1 pandemic influenza among non-medical personnel, medical students and medical professionals

To study the awareness of swine flu among general population, medical students and medical professionals Descriptive study. Place and Duration of Study: Study was carried out in the department of Pathology [Microbiology], Army Medical College, the Military Hospital [MH] and Combined Military Hospital, Rawalpindi, from 17th January 2010 to 24th January 2010. Materials and A total of 519 people were included in this study, 228 were medical students, 181 were doctors and 110 were non health care professionals. A questionnaire with closed questions regarding the awareness of the basics of swine flu was required to be filled by all participants. It was found that the general population was totally unaware to the extent that 1/3rd had never heard of this disease. The survey of medical students showed reasonable knowledge in the senior classes [80-85%] knew the main features of the disease], whereas the 2nd year students had the awareness around 40-45%. Among the medical professionals the knowledge was excellent in the seniors, whereas the trainees and house officers had suboptimal knowledge of swine flu. The result of this study show a significant gap between the seniors specialists and the juniors and this gap warrants immediate bridging. The general public also need to be informed by various means more comprehensively

Extended-spectrum beta-lactamase production in sbigella flexneri

Emergence of multidrug-resistant strains of Shigella is a growing concern across the globe. Third-generation cephalosporins are used for treating infections caused by multidrug-resistant Shigellae. However, resistance to these cephalosporin antibiotics due to extended-spectrum beta-lactamases, has emerged as a new problem. So far extended-spectrum beta-lactamases producing Shigella has not been reported from Pakistan. We report such a case in Shigella flexneri from an 8-year old girl with acute dysentery

Susceptibility pattern and frequency of extended specturm beta-lactamase producers in a tertiary care hospital

Infections with extended spectrum beta-lactamase [ESBL] producing organisms continue to be associated with higher rates of mortality, morbidity and health care costs. This study was carried out to find out frequency and sensitivity pattern of ESBL producers among Gram negative rods [GNRs] from clinical isolates of a tertiary care Hospital. A total of 1430 GNRs were recovered from 2347 clinical samples received from admitted patients in Military Hospital Rawalpindi. All samples were dealt by standard microbiological methods. Extended spectrum beta-lactamase detection in these isolates was done by double disc approximation test of Jarlier et al, 1988. Frequency of GNRs among clinical isolates was 61% and about 33% of these were ESBL producers. Escherichia coli were the most frequent ESBL producers followed by Klebsiella pneumoniae. Most of the ESBL producers were isolated from urine followed by catheter tips and pus. Among all the antibiotics tested, ESBL producers showed highest susceptibility to carbapenems followed by amikacin. Organisms showing resistance pattern similar to ESBLs were also significant. Continued surveillance by clinical microbiology laboratories, judicious use of antimicrobial agents and implementation of infection control measures are recommended to reduce the frequency of ESBL isolates

Frequency of diabetes mellitus, viral hepatitis among cataract patients in a Saudi military hospital

To determine the frequency of diabetes mellitus and viral hepatitis B and C among cataract patients of Saudi origin. Record of ninety nine patients of Saudi origin who were operated for age related cataract was analyzed to note frequency of Diabetes mellitus and viral hepatitis B and C. This was besides the mandatory lab screening for HIV and sickle cell disease. Twenty nine percent patients were diabetics. Ten patients were reactive to viral hepatitis screening with six positive for hepatitis B surface antigen and four positive to anti Hepatitis C virus antibodies. Pre-op screening for common diseases should be mandatory for safety of patients and medical staff