Objectives: Metallo-beta lactamase (MBL) producing non fermenter Gram negative bacilli is an emerging warning and cause of worry as they have established as one of the most feared resistance mechanisms and are the foremost cause of nosocomial infections worldwide. Carbapenem, including Imipenem, Meropenem and Doripenem are often used as a last remedy for treatment of infections caused by Pseudomonas aeruginosa, Acinetobacter and other Gram-negative. The present study was designed to explore the distribution of imipenem resistant non-fermenter Gram-negative bacilli isolates in different age groups. Study Design: Cross sectional Descriptive study. Setting: Microbiology laboratory, PGMI, Lahore. Period: January 2015 to December 2015. Material & Methods: 53 imipenem resistant NFGNB that were isolated from appropriate sampling of patients suffering from several infections were analyzed by using different standard microbiological techniques like microscopy, culture methods, biochemical reactions and antibiotic susceptibility using Kirby-Bauer method. MBL recognition was performed by imipenem-2MPA double disc synergy test (DDST). Results: This study shows the frequency of imipenem resistant non-fermenter Gram-negative bacilli isolated from various clinical wards. Maximum NFGNB were recovered from surgery/surgical allied 35.84% followed by ICU 28.3%, medicine /medicine allied 20.75%, pediatrics 9.4% and gynae/ obs 5.6% respectively. MBL production was identified among different imipenem resistant non-fermenter Gram-negative bacilli isolates by DDST using 2-MPA. Out of total 53 imipenem resistant non-fermenter Gram Negative Bacilli 37 Pseudomonas aeruginosa 20(54.05%) were MBL positive. Out of 13 Acinetobacter baumannii and 2 Pseudomonas luteola, 11(84.61%) Acinetobacter baumannii and 1(50%) Pseudomonas luteola were positive for MBL production. None of the Acinetobacter junii indicated MBL production. Conclusion: Double disc synergy test is operational for detection of MBL producers among NFGNB. It can be established in our routine clinical microbiology laboratories, for the MBL recognition especially in imipenem resistant isolates as of its cost efficiency.

Background and Objective: Dermatophyte infections require laboratory diagnosis before treatment is started. Although direct microscopy is routinely performed but culture of dermatophytes is the gold standard. However, it takes about 4 weeks for species identification on primary media. Our aim was to compare dermatophyte test medium (DTM) as a screening medium for the isolation of dermatophytes in comparison with sabouraud dextrose agar (SDA). Methods: It was a comparative study carried out at the Department of Microbiology of Post Graduate Medical Institute, Lahore over a period of nine months. Samples were collected from one hundred patients with clinically suspected dermatophytoses after taking informed written consent. The samples were examined microscopically and then inoculated on two types of culture media, one Sabouraud dextrose agar (SDA) with added chloramphenicol, gentacin and cycloheximide and other dermatophyte test medium (DTM) with added chlortetracycline, gentacin and cyclohexamide. Results: Fungal growth was observed in fifty-six samples on culture. Out of the fifty-six positive on cultures, nineteen were that of dermatophytes. Out of n = 100 patients, ten were positive on SDA while n = 14 dermatophyte species were able to grow on DTM. A significantly higher positivity (P ³ 0.05) for isolating dermatophytes was observed by DTM as compared to SDA. DTM was able to isolate (71%) of the dermatophytes in first 10 days. Isolation rate of dermatophyte species was higher (73.68%) on DTM as compared to SDA which was 52.6%. Conclusion: Authors recommend the use of dermatophyte test medium for the primary isolation and identification of dermatophyte species to be more effective and time saving.

Antimicrobial resistance (AMR) is a devastating question that is threatening the health globally. The extensive and indiscriminative use of antibiotics has evolved a notorious resistance in Staphylococcus aureus.  This resistance developed through possession of mecA gene, which codes for modified penicillin binding protein (PBP2a) and the emergent strain being labeled “methicillin resistant Staphylococcus aureus”. Conventional phenotypic techniques for detection of MRSA rely on standardization of cultural characteristics. The drawbacks of diagnostic error to report MRSA include: poor prognosis, expensive treatment, dissemination of multi-drug resistant strains and even treatment failure. Latex agglutination method can be adopted as a more accurate and quick strategy for rapid detection of methicillin resistance. Objectives: To compare detection of mecA gene in methicillin resistant isolates of Staphylococcus aureus by latex agglutination and PCR; by assessing the sensitivity and specificity of both methods. Study Design: Descriptive Cross-Sectional study. Setting: Pathology Department, Post Graduate Medical Institute, Lahore. Period: From January 2015 to December 2015; according to standard operating procedures at Microbiology laboratory. Material & Methods: A total 713 consecutive, non-duplicate isolates of Staphylococcus aureus were processed. Methicillin resistance was determined using cefoxitin (30mg) by Kirby-Bauer method using CLSI guideline (2016), latex agglutination method; and PCR for mecA gene. Results: The results showed that out of 713 Staphylococcus aureus isolates, 92 (12.90%) isolates were resistant to cefoxitin and were labelled as MRSA. majority MRSA isolates recovered from pus (44.57%) and wound swab (20.65%), followed by blood (13.04%), fluid (8.70%), CSF (4.35%), CVP (3.26%), HVS (3.26%) and tracheal secretion (2.17%). By latex agglutination method, 87 (94.50%) were positive for PBP2a; while on PCR mecA gene was detected only in 82 (89.10%) MRSA isolates. When assessed with PCR (gold standard) the sensitivity and diagnostic accuracy of latex agglutination was 100% and 94.57%, respectively. Conclusion: Latex agglutination test can be employed as rapid and reliable diagnostic technique in MRSA isolates for mecA gene detection, where resources for molecular methods are inadequate. This can effectually lessen the misdiagnosis of resistant strains, and over/ ill-use of antibiotics.

Objective: To determine the frequency and SCCmec type of nasally carried MRSA in HCWs of a tertiary care hospital. Methods: Nasal swabs were collected from three hundred and eighty healthcare workers working in various clinical wards of Lahore General Hospital, Lahore. The phenotypic resistance to methicillin was determined using Cefoxitin disk 30 µg. All the isolates showing Cefoxitin resistance were confirmed for mecA gene and typed for SCCmec I, II, III, IV (a, b, c, d) and V by PCR. DNA sequencing was done for random isolates for all the SCCmec types recovered in the present study. Results: Out of 380 nasal samples, 89 (23.42%) cultures yielded the growth of S. aureus out of which 31 (34.83%) were MRSA. The overall frequency of MRSA among all the HCWs was 8.2%. Overall, 47 SCCmec elements were found in total 29 MRSA isolates. Out of 29 MRSA isolates, 13 (44.82%) were hospital acquired, 7 (24.13%) were community acquired and 9 (31.03%) isolates had SCCmec types of both hospital acquired and community acquired origins. Conclusion: The colonized healthcare workers are harboring Hospital-acquired MRSA more frequently and can act as being acting as mixing bowls of different SCCmec genes. Our study emphasizes the need for the formulation of regular nasal decolonization policies for effective infection control within our healthcare setups.

Aim: To compare the resistance amongst Gram negative bacteria against imipenem and meropenem. Study Design: Prospective, non-randomized, descriptive study. Place and Duration of Study: Department of Microbiology, Mughal Laboratories, Lahore from 1stJuly 2019 to 31stDecember 2019. Methodology: One hundred culture samples received, bacteria isolated and their susceptibilities to imipenem and meropenem were compared. Organisms were recognized by the microbiological techniques according to the current standards and susceptibility testing was done according to the guidelines of Clinical and Laboratory Standards Institute (CLSI) 2020by using Kirby Bauer Disc diffusion method. Results: Salmonella typhi, Citrobacter species and Proteus species were 100% sensitive to imipenem. The rest of bacterial isolates had sensitivities to E. coli 88%, Acinetobacter 80%, Klebsiella species 67% and Peudomonas species 64%. The meropenem is highly resistant in all the bacteria as compared to imipenem. Conclusion: Increasing the trend of carbapenem resistance amongst Gram negative bacteria excluding Salmonella typhi was recorded. Key words: Gram negative rods, Resistance, Spectrum