Mutational analysis of HCV gene encoding E1 glycoprotein

Objective: to evaluate the variations in HCV glycoprotein E1 gene and to map epitopes in the variable E 1 regions informing the development of an effective vaccine against HCV

Methods: to isolate the E1 gene, RNA extraction was done by using the kit method and then it was converted to the cDNA. Confirmation of HCV presence in the collected samples was done through highly conserved core primers. This was then followed by PCR amplification for E1 gene. The sequenced E 1 genes were translated in silica into protein sequences

Results: a these proteins sequences were then analyzed for the presence of B-cell and T-cell epitopes; two B-cell epitopes [CSLYPGHLSGHRMAWD, TASIRSHVDLLVGMT] and one T-cell epitope [QAFTFRPRR] were found useful. These could be helpful in the formation of a proper vaccine against HCV

Conclusion: we found 2 B-cell epitopes and 1 T-cell epitope conserved in 3a genotype that may help in vaccine development

Trend of increased resistance to antibiotics in intensive care unit isolates of a tertiary care hospital over two years 2009 and 2010

To asses the antibiotic resistance pattern of Intensive Care Unit bacterial isolates over two year period; 2009 and 2010. This observational study was carried out on Intensive Care Unit isolates of Services Hospital Lahore in the Microbiology section of Department of Pathology, Services Institute of Medical Sciences Lahore. All samples processed for microbial cultures were tested for anti-biotic sensitivity / resistance pattern studied and compared. In 2009,790 samples and in 2010 886 samples were submitted from ICUs to Microbiology Section for culture. Of these, 42% and 46% were culture positive respectively. Gram negative isolates were 294 in 2009 and 308 in 2010. Resistance to all drugs tested was exhibited by 26 [8.78%] and 39 [12.60%] isolates in 2009, 2010. The total number of Acinetobacter isolated increased to 102 in the year 2010 from 74 in the year 2009 with 28 more Acinetobacters than in 2009 and the number exhibiting extensive drug resistance doubling to 28 from 14. Resistance to Imipenem, Tazobactem and Amikin drugs increased in Acinetobacters, Klebsiella and resistance of E coli to Imipenem also increased but decreased in Pseudomonas and E coli. ORSA and coagulase negative staphylococci with Oxacillin resistance were also on the rise, doubling in number from 12 to 25 and 14 to 31 in 2009 and 2010. Acinetobacter species are on the rise in the intensive care units as is their extensive and multi drug resistance pattern. Increasing Carbepenem resistance is alarming limiting our therapeutic options. Judicious use of antibiotics and curtailing nosocomial infections would deter this upward trend

Association of blood group types to Hepatitis B and Hepatitis C virus infection

Frequency of Hepatitis B Surface Antigen [HBsAg] and Hepatitis C Virus Antibodies [Anti-HCV] among blood donors of Lahore and their association with blood group types. To study the frequency of Hepatitis B Surface Antigen [HBsAg] and Hepatitis C Virus Antibodies [Anti-HCV] in blood donors of Lahore and to assess the association with blood group types. The design of study will be cross sectional descriptive study. It was held in the Pathology Department, Nawaz Sharif Social Security Hospital, Lahore, during the period January, 2006 to December, 2008. A total of 16695 blood donors were screened for HBsAg and Anti-HCV by rapid test devices based on immuno-chromatographic technique following the instruction given by the manufacturer. In the present study, devices manufactured by Acon, USA were used. The specimens reactive on screening by devices were confirmed on ELISA. The results were subjected to chi-square analysis for determination of statistical difference between the values among different categories. Among 16695 blood donors, 467 [2.79%] were positive for HBsAg and 1326 [7.94%] were positive for Anti-HCV. The frequency of HBsAg was seen to decrease significantly [p < 0.01] from 2006 to 2008 [4.23% to 2.31%]. However, frequency of anti-HCV was seen to rise significantly [< 0.01] from 2006 [6.69%] to 2008 [7.82%]. Comparison of HBsAg and anti-HCV positivity among RhD positive and RhD negative donors showed that there was no significant difference for HBsAg positivity [2.79% vs 2.85%]. However, significantly higher number of RhD positive donors had HCV infection as compared to RhD negative donors [8.25% vs 3.66%]. High frequency of HCV infection in blood donors need implementation of strict screening policy for donors and public awareness campaigns about preventive measure to reduce the spread of this infection as well as other transfusion transmissible infections. Association of HCV infection with blood group types needs more studies to get more knowledge about this aspect

Children with acute viral hepatitis: a one year study at Shaikh Zayed Hospital, Lahore

Objectives: To study the clinical spectrum, outcome and etiological agents of acute viral hepatitis in admitted patients at Paediatric Department Shaikh Zaycd hospital, Lahore. Setting: Paediatric Department Shaikh Zayed hospital, Lahore

Duration: One Year from November 2004-0ctober 2005. Study design: Retrospective Descriptive study. Inclusion criteria: I. Age between 1-15 years; 2. All patients admitted with acute viral hepatitis in the Paediatric department. Exclusion criteria: 1. Children with drug induced hepatitis; 2. Children with chronic liver disease; 3. Children with obstructive jaundice

Results: A total of 69 patients out of 2391 admitted patients were included during that period out of them 43[62%] were males and 26 [38%] were females. In our study jaundice was the presenting complaint in 52[75%] patients, followed by vomiting in 48[69%], fever in 42[60%], dark colour urine in 30 [43%], anorexia in 25 [36%], abdominal pain in 24 [35%], yellow colour stool in 7[10%] and irritability in 2 [3%] patients. 13 patients had a positive family history of hepatitis out of them 3[4.3%] had Hepatitis A virus [HAY], 7[10%] Hepatitis B virus [HBY], and 3 [4.3%] had Hepatitis C virus [HCY]. History of blood transfusion was found in 7[10%] patients while it was negative in remaining 62[90%]. Only 2[2.8%] patients were vaccinated against HAY and 9 [13%] against HBY. Icterus was present in 52[75%], 65[94%] had tenderness in the right hypochondrium, hepatomegaly in 57 [83%] and splenomegaly in 7% and one patient was drowsy at the time of admission. All the patients were screened against HAY, HBY, HCY, HOV and HEY in a serial wise approach. Out of them 50[72%] patients were HAY IgM positive, 4[5.7%] HbsAg positive, 2[2.8%] had anti-HCY, 2[2.8%] had HEY lgM and 1[1.4%] had HDY+HbsAg. In 10 [14%] patients no viral marker was detected out of these five viruses. Out of 69 patients 4[5.7%] developed hepatic encephalopathy. Out of these 2 pts. expired and 2 survived. A total of 67[97%] were discharged and 2 [3%] expired

Conclusions: Jaundice was the main presenting complaint followed by vomiting with significant recovery and Hepatitis A was the· most common cause of acute viral hepatitis in children admitted in hospital

Seroprevalence of rubella IGG and IGM antibodies in infants suspected of having rubella infection

This study was conducted to assess clinical and laboratory screening for suspected congenital rubella syndrome [CRS] and to evaluate the efficacy of reporting of CRS cases at the Children's Hospital and the Institute of Child Health. A total of 100 infants suspected of having rubella infection were included in the study. Rubella titer was used as an indicator. Enzyme linked immunosorbent assay was performed to detect rubella specific IgM and IgG. The data was analyzed on infants 'birth weight, infants' clinical characteristics and laboratory evaluation. The study shows that out of 100 Infants, only three [3%] met the definition for confirmed rubella whereas thirty-four patients were positive for Rubella IgG only. Ninety-seven other children had clinical presentation that met the definition for a probable case but when they were tested for Rubella specific IgM antibodies, the tests were negative. Rubella IgG positive patients were further categorized on the basis of their age; 33 [97%] infants were of less than six months and only 1 [3%] infant was of more than six months. Average age of the patients was 2.2 months [SD +/- 1.55]. The common clinical presentations in rubella positive patients [n=3] were failure to thrive [100%], cataract [67%], patent ductus arteriosus [67%], microcephaly [67%], intracranial calcification [33%], buphthalamus [33%], and hepatosplenomegaly [67%]. With regard to hematological abnormalities in the three rubella confirmed cases, three had anemia [loon two infants had thrombocytopenia [67%], and only one infant had leucopenia [33.3%]. The relatively high rate of susceptibility indicated a risk of a rubella outbreak, and the resulting. Congenital rubella syndrome is an under-recognized public health problem in Pakistan and can be reduced by vaccinating all seronegative women. There is an urgent need for the collection of appropriate data to estimate the cost effectiveness of a potential Rubella Control Programme

Staphylococcus aureus and MRSA nasal carriage in general population

Assessment of Staphylococcus aureus [S. aureus] and MRSA nasal carriage in our general population. Cross-sectional analytical study. A private Pathology Laboratory in Lahore city from January 2002 to December 2003. Nasal swabs were obtained from the apparently healthy persons accompanying the patients attending the laboratory. These swabs were inoculated on blood and mannitol salt agar plates and incubated at 37°C. The isolate[s] were identified as S. aureus on the basis of colony morphology, Gram staining, catalase and coagulase tests. Sensitivity to Oxacillin was determined using standard Kirby Bauer technique. Nasal swabs from 1660 subjects from the community were studied. Out of these samples, a total of 246 [14.82%] samples were positive for growth of S. aureus [nasal carriers of S. aureus]. Out of 246 S. aureus isolates, 48 [19.51%] isolates were methicillin resistant S. aureus [MRSA]. Nasal carriage was higher in males [15.47%] as compared to females [13.26%], in urban areas [16.99%] as compared to rural areas [11.32%] and in the year 2002 [16.02%] as compared to year 2003 [13.08%]. However, the difference was significant only for the urban vs. rural comparison [p<0.05]. MRSA isolates were also significantly higher among S. aureus isolates from the nasal cavities of urban subjects [22.98%] as compared to rural ones [11.11%]. Maximum nasal carriage was present in the age group upto 9 years [20.23%] with decrease in the age groups 10-19, 20-29 and 30-39 years followed by small rise in the older subjects. The nasal S. aureus carriage as well as methicillin resistance among these isolates are more common in urban community