Comparison of dried blood spots with conventional blood sampling for diagnosis of hepatitis b & c through serological and molecular technique; a pilot study.

OBJECTIVE: To compare the sensitivity and specificity of dried blood spot with conventional blood sampling for serological and molecular testing of hepatitis B and C viruses. METHODS: The case-control study was conducted at the Pakistan Health Research Council Specialised Centre for Gastroenterology and Hepatology, Karachi, from May,2015 to April,2016 and comprised patients who were hepatitis B surface antigen-positive (group 1), anti-hepatitis C virus positive (group 2), hepatitis B virus deoxyribonucleic acid-positive (group 3), and hepatitis C virus ribonucleic acid-positive (group 4). A group of controls had healthy subjects negative for both hepatitis B and C viruses. Blood samples were collected using the conventional as well as the dried blood spot method using finger prick. Relevant tests were run for each subject using both the samples at baseline and after 3 and 6 months of storage. Receiver operative characteristic curve was plotted to determine the ideal cut-off points for dried blood spot testing and corresponding sensitivity and specificity. Data was analysed using SPSS 19. RESULTS: Of the 100 subjects, there were 20(20%) in each of the four patient groups and 20(20%) in the control group. Sensitivity of dried blood spot method was 95.2%, 95%, 80% and 70% for groups 2, 1, 4 and 3 respectively when tested within a week of sampling. Specificity was 100% for all the four groups. There was a significant correlation of the two methods for all the four parameters tested (p<0.01). CONCLUSIONS: Dried blood spot sampling correlated well with the conventional blood sampling method for serological and molecular testing.

Report - Colonization rates and immune status of children against Streptococcus pneumoniae before the introduction of pneumococcal conjugate vaccine.

Pneumococcal infections contribute to high morbidity and mortality in Pakistan. Pneumococcal colonization rates and immune status of children before and after vaccination can serve as an important indicator to determine the success of immunization program. Aim of this study was to determine the pneumococcal colonization rate and immune status of children (<3-years) before the introduction of Pneumococcal immunization in EPI program. A cross sectional study of 3-months duration was conducted in collaboration with EPI Centre of NICH, Karachi. A total of 192 healthy children aged 3-months to 3-years were included in the study. Children with chronic or recurrent pulmonary disease were excluded. Demographic data and presence of any risks for pneumococcal colonization or immunity was recorded. Nasopharyngeal swabs were collected to determine the presence of S. pneumoniae and sensitivity pattern of the isolates. Blood samples were collected to determine immune status of these children. Sixty (31%) children were found to carry S. pneumoniae in their nasopharynx. Resistance to cotrimoxazole, erythromycin, chloramphenicol, penicillin and vancomycin was 100%, 30%, 25%, 22% and 0% respectively. Mean anti-PCP titre of children was 23.3mg/L. Mean anti-PCP titre in children who had previously suffered from RTI was significantly increased (p-value 0.039). This study may serve as baseline report for evaluating the success of pneumococcal immunization in future. A significant drop in nasopharyngeal colonization rates post vaccination would depict the success of pneumococcal immunization.

Emerging trends of antimicrobial resistance in Helicobacter pylori isolates obtained from Pakistani patients: The need for consideration of amoxicillin and clarithromycin.

OBJECTIVE: To present antimicrobial susceptibility profile of helicobacter pylori isolated from gastric biopsies of dyspeptic patients. METHODS: The cross-sectional study was conducted from March 2008 to May 2013 at the Jinnah Postgraduate Medical Centre (JPMC), Karachi, and comprised patients with suspected peptic ulcer disease who underwent upper gastrointestinal endoscopy. Multiple gastric antral biopsies were taken. One sample was imbedded in the Campylobacter-like organism test, while other biopsies were processed for culture and confirmed on polymerase chain reaction. Isolates were subjected to antimicrobial susceptibility testing and minimum inhibitory concentrations of antimicrobial agents were estimated by Epsilometer test. SPSS 21 was used for statistical analysis. RESULTS: Of the 889 samples available, 254(28.6%) were positive after campylo-like organism test, of them, 92(36.2%) isolates were cultured. Susceptibility test showed 90(97.8%) resistance to metronidazole, 25(30.1%) ofloxacin, 6(16.2%) levofloxacin, 14(15.2%) furazolidone, 5(5.4%) clarithromycin, 4(4.3%) tetracycline and 2(2.2%) isolates showed resistance to amoxicillin. Resistance to more than one drug was observed in 37(40.2%) isolates. CONCLUSIONS: Low clarithromycin resistance was surprising since Pakistan is considered a hot bed of drug-resistant bacteria.

Is chlorhexidine-gluconate superior than Povidone-Iodine in preventing surgical site infections? A multicenter study.

OBJECTIVE: To compare the efficacy of povidone-iodine and chlorhexidine gluconate scrubs in preventing surgical site infections. METHODS: The randomised controlled clinical trial was conducted from May 2012 to April 2013 in two public-sector hospitals of Pakistan; one each in Karachi and Islamabad. Patients undergoing clean or clean contaminated surgeries were included and were randomly assigned to one of the two groups: group I comprised patients whose skin was preoperatively disinfected using 10% povidone-iodine, and in group II by 2% chlorhexidine gluconate in 70% alcohol. A predesigned proforma was filled for all patients to record demographic data, diagnosis, surgical procedure and antibiotic used. Patients in both groups were followed up for one month postoperatively to monitor any signs of surgical site infections. SPSS 16 was used for statistical analysis. RESULTS: Of the 388 patients from the two hospitals, 220(57%) were in group I and 168(43%) were in group II. Surgical site infection was observed in 22(10%) cases in group I and 12(7.1%) in group II (p=0.324). Pseudomonas aeruginosa (23.5%) was the predominant pathogen associated with surgical site infections followed by Staphylococcus aureus (17.6%). CONCLUSIONS: Chlorhexidine gluconate was associated with lower infection rates compared to povidone-iodine; but it was not statistically significant.

Seasonal Trends with the Comparison of Venous and Capillary Blood for Diagnosis of Malaria among Children in Karachi, Pakistan.

Background: More than 250 million people are infected by malaria parasites annually while around one million children less than 5 years of age die every year due to malaria. We aimed to assess the seasonal trends and usefulness of capillary and venous blood for rapid diagnosis of malaria. Methods: This cross-sectional study of 18 months duration was conducted at the National Institute of Child Health (NICH), Karachi. All patients reporting fever as chief complaint were recruited as study subjects. A semi-structured questionnaire was used to collect demographic information, presenting complaints, awareness of caregivers regarding malaria, preventive measures and history of malaria fever. Three ml Venous (2-3ml) as well as peripheral blood (3-4 drops) samples of all patients were collected for microscopy and rapid diagnostic tests (RDTs). Results: Out of total 477 patients with fever Venous and Capillary Blood RDTs methods detected 33(6.9%) and 30(6.3%) as the malaria positive while Venous and Capillary Blood Microscopy detected 30(6.1%) and 32(6.7%) cases respectively. Plasmodium Vivax infection was the most prevalent (87.9%) and majority (39.39%) of the cases occurred in the quarter, July to September. Conclusion: July to September is the peak season for malaria and P. Vivax (87.9%) is the predominant strain in Karachi. Venous and capillary blood are equally useful for malaria diagnosis however, convenience and less invasiveness may justify the preference of capillary blood over venous blood for early diagnosis of malaria.

Use of the Health Belief Model for the Assessment of Public Knowledge and Household Preventive Practices in Karachi, Pakistan, a Dengue-Endemic City.

BACKGROUND: Prevention is most effective in reducing dengue infection risk, especially in endemic countries like Pakistan. Evaluation of public awareness and health beliefs regarding dengue fever (DF) is important for devising disease control strategies. This study assessed dengue knowledge, health beliefs, and preventive practices against DF in different socioeconomic groups of Karachi, Pakistan. METHODOLOGY: In this community-based cross-sectional study, 6 randomly selected towns were visited, 2 persons (man and woman) per household were interviewed using a structured questionnaire, and household practices were observed. Information regarding DF was shared through a printed pamphlet. Multivariate logistic regression analysis of variables associated with dengue knowledge and practices was conducted. PRINCIPAL FINDINGS: We interviewed 608 Karachi residents (mean age: 33.2 ± 13.35 years); 7.7%, 71.9%, and 20.4% had a high, middle, and low socioeconomic status, respectively. The mean knowledge score was 6.4 ± 2.10 out of 14. The mean preventive practices score was 9 ± 1.8 out of 17. Predictors of dengue knowledge were perceived threat (odds ratio [OR] = 1.802; 95% confidence interval [CI] = 1.19-2.71; p = 0.005), self-efficacy (OR = 2.910; 95% CI = 1.77-4.76; p = 0.000), and television as an information source (OR = 3.202; 95% CI = 1.97-5.17; p = 0.000). Predictors of dengue preventive practices were perceived threat (OR = 1.502; 95% CI = 1.02-2.19; p = 0.036), self-efficacy (OR = 1.982; 95% CI = 1.34-2.91; p = 0.000), and dengue knowledge (OR = 1.581; 95% CI = 1.05-2.37; p = 0.028). CONCLUSIONS: Public knowledge about DF is low in Karachi. Knowledge, threat perception, and self-efficacy are significant predictors of adequate dengue preventive practices. Prevention and control strategies should focus on raising awareness about dengue contraction risk and severity through television. Health messages should be designed to increase individual self-efficacy.

High prevalence of typhoidal Salmonella enterica serovars excreting food handlers in Karachi-Pakistan: a probable factor for regional typhoid endemicity.

BACKGROUND: Typhoid fever is the persistent cause of morbidity worldwide. Salmonella enterica serovar's carriers among food handlers have the potential to disseminate this infection on large scale in the community. The purpose of this study was to determine the prevalence of typhoidal S. enterica serovars among food handlers of Karachi. METHODS: This cross-sectional study was conducted in Karachi metropolis. A total of 220 food handlers were recruited on the basis of inclusion criteria from famous food streets of randomly selected five towns of Karachi. Three consecutive stool samples were collected from each food handler in Carry Blair transport media. Culture, biochemical identification, serotyping, and antimicrobial susceptibility tests for S. enterica serovars were done. RESULTS: Out of 220 food handlers, 209 consented to participate, and among them, 19 (9.1%) were positive for S. enterica serovars. Serotyping of these isolates showed that 9 (4.3%) were typhoidal S. serovars while 10 (4.7%) were non-typhoidal S. serovars. Of the typhoidal S. serovars, 7 were S. enterica serovar Typhi and 1 each of S. enterica serovar Paratyphi A and B. The resistance pattern of these isolates showed that 77.7% were resistant to ampicillin and 11.1% to cotrimoxazole. All typhoidal S. enterica serovar isolates were sensitive to chloramphenicol, ceftriaxone, cefixime, nalidixic acid, and ofloxacin. CONCLUSIONS: Carrier rate of typhoidal S. enterica serovars in food handlers working in different food streets of Karachi is very high. These food handlers might be contributing to the high endemicity of typhoid fever in Karachi, Pakistan.