Objective: To determine incidence of MRSA in our community and its antibiotic susceptibility pattern. Study Design: Cross Sectional study. Setting: Department of Pathology, Sahara Medical College Narowal. Period: January 2020 to June 2020. Material & Methods: Bacterial isolates were taken from the samples of blood, pus and other body fluids sent to the hospital laboratory to determine culture and sensitivity pattern. Those samples positive for staphylococcus aureus were included in the study for further examination. MRSA were detected using conventional technique (catalase, coagulase and DNase methods) and confirmed by phenotypic and molecular characterization techniques (cefoxitin and oxacillin sensitivity, mec-A gene and 16S rRNA genes sequencing methods). Their antibiotic sensitivity was determined using standard Kirby Bauer disc diffusion technique. Chi square test was applied on collected data using SPSS-24. Results: Total 158 bacterial isolates containing staphylococcus aureus were included in the study. MRSA (multi drug resistant staphylococcus aureus) was detected in 33.5% samples and MSSA (multi drug sensitive staphylococcus aureus) was detected in 66.5% samples. Frequency of MSSA was more than MRSA. Male gender was more affected (60%) than female (40%). All bacterial isolates containing staphylococcus aureus were resistant to oxacillin and cefoxitin. MRSA isolates were sensitive to vancomycin, linezolid, teicoplanin, fosfomycin and fusidic acid, rifampicin, clindamycin, minocyclin and chloramphenicol, while resistant to penicillin, azithromycin, ciprofloxacin, erythromycin and co-trimoxazole. Conclusion: MRSA is detected by conventional technique followed by phenotypic and molecular characterization methods for confirmation. Methicillin sensitive staphylococcus aureus was detected more commonly than methicillin resistant staphylococcus aureus in our study sample. Proper culture and antibiotic sensitivity pattern is necessary to deal with MRSA.

Objective: To determine frequency of gram negative bacterial isolates and their antibiotic sensitivity pattern among patients with chronic suppurative otitis media. Study Design: Descriptive Cross Sectional study. Setting: Department of Pathology Sahara Medical College Narowal. Period: January 2020 to June 2020. Material & Methods: Total 126 bacterial isolates were taken from the patients having unilateral or bilateral ear discharge reporting to OPD or emergency department during study period. After taking detailed history, examination and baseline investigations diagnosis was confirmed. Specimens of ear discharge were taken from all patients and sent to the pathology department of study institution to determine culture and sensitivity pattern of Gram-negative bacterial isolates. Consent was taken from ethical review committee. Data was analyzed on SPSS-24. Chi square test was applied on collected data. Results: Bacterial isolates were taken from 126 cases having age 2-54 years with mean age of 11.45±5.73 years of either gender. Mostly children less than 8 years of age were found with otitis media. Gram positive bacteria were detected in 41.3% and gram negative in 55.6% and candida albicans in 3.2% out of total 126 bacterial cultures. Out of gram negative cultures, pseudomonas aeruginosa was found in 41.4%, proteus mirabilus in 44.3% and E.coli in 14.3% specimens. Conclusion: Gram negative bacteria are common cause of chronic suppurative otitis media in our region, mostly affecting young children. There is variable antibiotic sensitivity pattern depending on cultural, climate changes and previously taken antibiotics.

Objective: To determine frequency, molecular characterization and sensitivity of bacterial isolates against commonly used antibiotics in neonatal sepsis. Study Design: Cross Sectional study. Setting: Department of Pathology Sahara Medical College Narowal. Period: October 2019 to March 2020. Material & Methods: Neonates admitted in neonatal intensive care unit (NICU) of study institution having signs and symptoms of neonatal symptoms such as fever, irritability, seizures, anorexia and lethargy, were included in the study using consecutive sampling technique. Blood sample from all study patients taken and sent for culture to determine bacterial isolates and antibiotic sensitivity against commonly used antibiotics for neonatal sepsis. Bacterial isolates identification was done using standard bacteriological technique performed by modified Kirby & Bauer disc diffuse method as per Clinical and Laboratory standards institute (CLSI) guidelines. Results: Total 200 cases were studied having neonatal sepsis and admitted in NICU including 58% female and 42% male children. Bacterial growth occurred in 10.5% samples and in 89.5% samples no bacterial growth seen. There were 1% samples with gram positive and 9.5% samples with gram negative bacterial isolates. Klebsiella was the commonest organism isolated in 38.1% cases out of total positive isolates. There were 72.5% neonates having age 1-14 days and 27.5% neonates having age 15-28 days. Conclusion: Gram negative bacteria are common cause of neonatal sepsis, out of which Klebsiella is the commonest organism. Antimicrobial drug resistance in different infections is a serious emerging issue.

Objective: To determine frequency and pattern of extended spectrum beta lactamase in Klebsiella pneumoniae and E. coli in bacterial cultures. Study Design: Cross Sectional study. Setting: Department of Pathology Sahara Medical College Narowal. Period: January to June 2020. Material & Methods: Total 1100 bacterial isolates were studied out of them 655 E.coli and 445 of klebsiella pneumoniae. All samples were subjected to double disc diffusion method using third generation cephalosporin and amoxacillin-clavulanic acid for detection of ESBL. Data was analyzed using SPSS software version 24, and results were calculated in the form of frequency, percentage and standard deviation. P-value ≤0.05 was taken as statistically non-significant. Results: There were 48.9% male and 51.1% female subjects out of 1100 total cases. E.coli was detected in 59.5% and klebsiella in 40.5% samples. Total 44.2% samples were positive for ESBL enzyme. Of 655 E.coli samples 40.8% and of 445 Klebsiella pneumoniae samples 49.2% were positive for ESBL enzyme. Conclusion: It is necessary to detect ESBL positive Klebsiella pneumoniae and E.coli in laboratory workflow to avoid unnecessary use of antibiotics and development of resistance against them.

Introduction: The present study was intended to isolate, characterize and investigate the prevalence of virulence genes encoding fibronectin (fnbA and fnbB) adhesive molecules of S. aureus from dental patients attending outpatient department of THQ Hospital Pasur. Methodology: A total of 100 oral samples were collected from dental patients, pure cultures was segregated and identified through conventional microbiological methods to evaluate the prevalence of S. aureus. Isolates were further characterized by using specific primers for genotype fnbA and fnbB. Results: Results of the study declared that 68 samples were positive (68%) for staphylococcus aureus on the basis of growth on selective media and appearance of typical colonies supported by gram staining. These gram positive staphylococci were positive (86.7%) in coagulase and catalase testing. The results of polymerase chain reaction (PCR) revealed that 47 (69.1%) and 23 (33.8%) isolates showed amplification with type specific primer 23Sr RNA and NUC gene respectively. Furthermore, fnbA and fnbB type specific genes of S. aureus did not show any amplification in PCR reaction. Conclusion: Irrespectively the data in the present study showed the prevalence of S. aureus is significantly high in males and of age group of 20-40 years but no positive result was found for prevalence of fnbA and fnbB genes. All the S. aureus isolates were highly sensitive to vancomycin, linezolid and clindamycin.