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Objective: To determine the diagnostic accuracy of CHROMagar Orientation for its ability to support the growth and identification of uropathogens by keeping API 20E/20NE as a gold standard. Study Design: Cross-sectional study. Place and Duration of Study: Department of Microbiology, Armed Forces Institute of Pathology, Rawalpindi Pakistan, from Jun to Nov 2019. Methodology: A total of 470 midstream specimen of urine from patients suspected of UTIs were analyzed. Urine specimens were inoculated by 1µL calibrated sterilized loop each on CLED Agar, Blood agar and CHROMagar Orientation media plates. Culture plates were incubated at 35.00±2.00ºC for 18 to 24 hours. As per standard protocols, isolates were identified by colony morphology, Gram stain, catalase, oxidase, DNAase, coagulase, and API 20E/20NE. In addition, the appearance of microorganism on CHROMagar Orientation media were identified according to the manufacturer's instructions. Results: Out of 470 samples, 211 showed no growth, 90 were classified as mix growth (polymicrobial), and 169 samples were positive (unimicrobial) for the growth of uropathogens. All the samples positive for bacterial growth on conventional media were also positive on CHROMagar orientation and vice versa. The sensitivity and specificity of CHROMagar Orientation for differentiation of uropathogens were 99% and 100%, respectively. Conclusion: The overall findings of our study suggested that CHROMagar supports the growth of all uropathogens. It provided the rapid identification of organisms based on their colour morphology, even in a mixed bacterial culture, growth within 24 hours easily. [ABSTRACT FROM AUTHOR]

Objective: To determine maternal vaginal colonization with Group B Streptococcus using both recto-vaginal swab and mid-stream urine in order to determine which specimen type can give better results. Study Design: Cross-sectional study. Place and Duration of Study: Armed Forces Institute of Pathology, Rawalpindi Pakistan, Department of Gynecology and Obstetrics, Combined Military Hospital and Pak Emirates Military Hospital, Rawalpindi Pakistan, from Jan to Jun 2022. Methodology: Recto-vaginal swabs and mid-stream urine were taken from 194 pregnant women between 36th and 37th weeks of pregnancy. Swabs were inoculated on blood and MacConkey agar while urine specimen was inoculated on blood and CLED agar. All plates were incubated at 37°C in either ambient air or CO2 incubator. Identification of Group B Streptococcus was made on the basis of colony morphology (β-haemolytic colonies on blood agar), Gram stain, catalase test and confirmation were done by means of latex agglutination tests. Results: In comparison to urine, a Group B Streptococcus carriage rate of 8.2% was detected in recto-vaginal swab. Urine specimen of only 3 patients yielded growth of Group B Streptococcus giving a urine carriage rate of 1.5%. Conclusion: Recto-vaginal swab gives a better insight to the presence of Group B Streptococcus as compared to mid-stream urine. [ABSTRACT FROM AUTHOR]

Objective: To determine the frequency of respiratory syncytial virus (RSV) among hospitalized children with acute lower respiratory tract infections (ALRI) and associated risk factors. Study Design: Comparative cross-sectional study. Place and Duration of Study: Microbiology Department, Armed Forces Institute of Pathology, Rawalpindi Pakistan, from Aug 2018 to Feb 2019. Methodology: Gram staining and culture inoculation of all specimens were performed per recommended guidelines. The Gram stain results on day-0 were compared with the results of the culture on day-2. There was no discrepancy if similar organisms were obtained on culture as on Gram stain. Nevertheless, if the Gram stain and culture results were not similar, it was termed a discrepancy. The consultant microbiologist reviewed all discrepant slides, and if not resolved, possible causes of error were sought, and the results documented. Results: Of the total 300 clinical specimens, errors were observed in the initial gram staining of 29 specimens (9.7%), whereas 271(90.3%) specimens were error-free. Upon evaluating these 29 errors, 11(38.0%) were observer errors which were resolved when reviewed by a consultant microbiologist. 14(48.0%) were technical errors, and 4(14.0%) results were discrepant due to the presence of anaerobic organisms, missed on initial aerobic cultures. Conclusion: The frequency of Gram stain errors in our study (9.7%) is not very high; nevertheless, it can have severe consequences in critical samples from seriously ill patients if wrong empirical antimicrobial treatment is begun based on a wrong initial Gram stain result. [ABSTRACT FROM AUTHOR]

Objective: To determine the molecular diversification and frequency of mecA Gene in MRSA infections of hospitalised patients at Islamabad, Pakistan. Study design: Cross-sectional study. Place and Duration: Pakistan Institute of Medical Science, Islamabad, Pakistan, from Sep 2017 to Sep 2018. Methodology: To explore the occurrence of pathogenic S. aureus in the hospitalised patients of Islamabad, 500 samples were collected. All the isolates were further characterised by gram staining, catalase test and DNAse media analysis validating the presence of S.aureus. The overall frequency of S.aureus was 19.5% in all the isolates. Six highly distinguished colonies were further evaluated through colony PCR by 16S r RNA analysis, and results revealed 99.9% homology of isolates with S.aureus. The degree of resistance was further evaluated for 12 isolates based on the previous MRSA resistance pattern. The extraction and concentration of DNA were done by the CTAB and Nanodrop methods. Then primers were designed for the presence of the mecA gene of S. aureus. Results: Results showed 51 (52.7%) MSSA and 45 (47.4%) MRSA among all the isolates. Colony PCR by 16S r RNA analysis results revealed 97 (99.9%) homology of isolates with S.aureus. PCR of all the twelve extracted DNAs of isolates showed the frequency of the mecA gene in all the isolates. Conclusion: The study is the first report the presence of resistance mecA gene in the hospitalised patients of Islamabad. It has depicted a resistant gene's presence and showed the rising trend of resistance in pathogenic S. aureus. [ABSTRACT FROM AUTHOR]

Objective: To determine the diagnostic accuracy of Colistin agar for detection of Colistin resistance in clinical isolates of Multi- Drug Resistant Gram-Negative Bacilli. Study Design: Cross-sectional validation study. Place and Duration of Study: Department of Microbiology, Armed Forces Institute of Pathology, Rawalpindi Pakistan, from Feb to Aug 2019. Methodology: A total of 100 Multi-Drug Resistant Gram-Negative Bacilli in clinical isolates were included. Isolates were identified using Gram stain, Catalase, Oxidase, API 20E, and API 20NE. After approval from the institutional ethical review committee, Colistin susceptibility was determined simultaneously by Colistin agar and Broth Micro Dilution Minimum Inhibitory Concentration method as per CLSI. For susceptibility criteria, EUCAST guidelines were followed. Results were validated with the gold standard test, i.e., Broth Micro Dilution. Results: Out of 100 Multi-Drug Resistant clinical isolates, the distribution was K. pneumoniae n=60, E.coli n=16, A. baumannii n=11, C. freundii n=8, and E. cloacae n=5. The sensitivity, specificity, positive predictive value, negative predictive value, and diagnostic accuracy of Colistin agar for detection of Colistin resistance, keeping Broth Micro Dilution Minimum Inhibitory Concentration method as the gold standard was 96.67%, 97.14%, 93.55%, 98.55%, and 97%, respectively. Conclusion: Colistin agar has excellent diagnostic accuracy for the detection of colistin resistance with standardized inoculum density. Due to its ease of use, cost-effectiveness, and accurate results, it can be used in lab setups deficient in manpower and advanced equipment for Broth Micro Dilution or genetic sequencing. [ABSTRACT FROM AUTHOR]

Objective: To assess the correlation between biofilm formation and azole antifungal susceptibility against plank tonic and sessile clinical isolates of C.albicans. Study Design: Prospective observational study. Place and Duration of Study: Combined Military Hospital Peshawar, from Jun 2016 to Sep 2017. Methodology: All standard microbiological procedures were carried out according to latest Clinical & laboratory standard institute (CLSI) guidelines. After gram staining and presumptive identification on CHRO Magar Candida, the isolates were biochemically identified by API AUX Candida as C.albicans. Planktonic antifungal susceptibility was carried out by Kirby Bauer disk diffusion method on 300 C.albicans isolates. Broth microdilution method was used to determine Minimum inhibitory concentration (MICs) of plank tonic cells and micro titer assay was used for assessment of biofilm formation by C.albicans. Results: In planktonic antifungal susceptibility, fluconazole was susceptible against 195 (65%) and voriconazole against 241 (80%) C. albicans isolates. C. albicans was found susceptible dose dependent (SDD) to fluconazole in 28 (9%) and to voriconazole in 21 (7%) isolates. Seventy-seven (26%) and 38 (13%) C.albicans isolates were found fluconazole and voriconazole resistant, respectively. Sessile antifungal susceptibility was carried out through broth micro dilution method in which 160 (53%) were susceptible, 42 (14%) were susceptible dose dependent SDD and 98 (33%) were resistance to voriconazole, and 161 (54%) were susceptible, 36 (12%) were SDD and 103 (34%) were found resistant to fluconazole. Biofilm forming isolates of C.albicans were observed to be 285 (95%).The p-value is highly significance i.e. <0.01 between the biofilm formation and azole antifungal susceptibility against plank tonic and sessile clinical isolates of C.albicans. Conclusion: Plank tonic C.albicans clinical isolates appeared more susceptible to voriconazole than fluconazole and sessile isolates. Biofilm formation was very high among all the isolates of C.albicans. [ABSTRACT FROM AUTHOR]

Objective: To investigate the prevalence of S. aureus in hospitalized patients of Islamabad. Study Design: Cross-sectional study. Study Duration: Pakistan Institute of Medical Science, Applied Microbiology and Biotechnology Lab, COMSATS Institute of Information Technology, Islamabad, from Sep 2017 to Sep 2018. Methodology: A total of 500 samples were collected. The isolates were divided into four study groups according to their source of origin i.e. group 1 (dermal group), group 2 (nasal group), group 3 (blood group) and group 4 (urine group). Gram staining, catalase test and DNA se media analysis were done for validation of S. aureus. Disc diffusion test (for antibiotic susceptibility), Oxacillin disc test (to differentiate between methicillin-resistant Staphylococcus aureus and methicillin-susceptible staphylococcus aureus) and minimal inhibitory concentration (for susceptibility to vancomycin), were performed. Results: Degree of the prevalence of staphylococcus aureus was 21%, 17%, 9% and 8% in group 1, 2, 3 & 4 respectively. The overall prevalence of staphylococcus aureus was 19.5% in all isolates. The disc diffusion test showed the descending resistance pattern of isolates i.e. 100, 94, 94, 76, 58, 55, 47, 43, 40 and 37% for penicillin, ciprofloxacin, Kanamycin, erythromycin, tetracycline, oxazolidinone, sulfamethoxazole, doxycycline, clindamycin, and cipoxin respectively. Minimal inhibitory concentration found only one sample resistant at 2ug/l concentration of Vancomycin. Moreover, Oxacillin disc test showed 52% methicillin-susceptible Staphylococcus aureus while 48.2% methicillin-resistant staphylococcus aureus among all isolates. Conclusion: There is an increase in the frequency of methicillin-resistant staphylococcus aureus. Single vancomycin resistant staphylococcus aureus strain was also isolated. [ABSTRACT FROM AUTHOR]

Objective: To determine in vitro efficacy of Imipenem/Relebactam against multi-drug resistant strains. Study Design: Cross-sectional study. Place and Duration of Study: Department of Microbiology, Fauji Foundation Hospital (FFH) Rawalpindi, Pakistan from Jul to Dec 2022. Methodology: All specimens from indoor and outdoor patients with infection due to Gram-negative organisms showing resistance to Imipenem disc (10µg) were included. Cultures were inoculated on Blood and MacConkey's agar. Imipenem-resistant Gram-negative species identification was made by Gram stain, Oxidase test and commercially available identification strips Analytical profile index 10S (API-10S) (from bioMerieux, Inc.).Imipenem-resistant species were identified, showing a zone diameter of less than 23mm against10µg Imipenem disc for Enterobacterale and Non-Enterobacterales, and for Pseudomonas species, zone size is ≤19mm. These resistant organisms were tested for sensitivity against Imipenem/Relebactam by E-strip (MTSTM -MIC Test Strip) (Liofilchem, Inc., Waltham, MA). Results: Out of 160 Carbapenem-resistant Enterobacterale and Non-Enterobacterales specimens, only 14 isolates (including 02 Pseudomonas spp.) showed susceptibility to Imipenem/Relebactam combination of drug. US-FDA Imipenem/Relebactam susceptibility interpretive criteria (STIC) was used. Conclusion: In our setup, only 8.75% isolates showed sensitivity to this newer drug combination Imipenem/Relebactum. This highlights the importance of evaluating newer drugs in our population before using them empirically. [ABSTRACT FROM AUTHOR]

Objective: To evaluate the burden of carbapenemases producers among carbapenem-resistant isolates. Study Design: Cross-sectional study. Place and Duration of Study: This cross-sectional study was conducted at microbiology lab, Pathology department, Allama Iqbal Medical College Lahore, from Jul 2016 to Apr 2017. Material and Methods: A total of 12126 clinical specimens were enrolled through a non-probability/consecutive sampling technique, every sample was processed for bacterial culture, identification was done on the basis of colonial morphology, Gram stain, and biochemical profile. All those isolates that were Gram-negative rods, were processed for carbapenemases detection by Modified Hodge test (MHT) in accordance with CLSI 2016. Results: Out of total n=12126 samples, culture positive were 35.9% (n=4361) of which 40.5% (n=1770) were Gram-negative rods, among these 9.6% (n=170) were carbapenem-resistant isolates of which 50% (n=85) were carbapanamases producers (MHT positive). Organisms wise carbapenem resistance was detected in Acineto- bacter species 44.%, Pseudomonas species 34%, E.coli 7%, Klebsiella species 8%, Proteus species 3%, Citrobacter species 1%, Enterobacter species 0.7% Among these isolates MHT positivity was as followed Acinetobacter species were 53%, Pseudomonas species 51%, E.coli 36%, Klebsiella species 50%, Proteus species 20%, Citrobacter species 66%, Enterobacter species 50%, department wise distribution of carbapenemases producer showed that among total sample received from surgical units 51.4%, ICU 65.3% medical units 43.5%, pediatric ward 71.4%, orthopedic 11.1% neurosurgery 20%, other wards 55.1% were MHT positive. The almost similar resistant pattern was observed in MHT positive and negative isolates. Conclusion: Emergence of carbapenem resistance is an alarming situation in clinical settings. [ABSTRACT FROM AUTHOR]

Objective: To determine the sensitivity of Carbapenem-resistant Pseudomonas aeruginosa (CARP) with the panel of antibiotics according to Clinical and Laboratory Standards Institute (CLSI) 2020 guidelines. Study Design: Cross-sectional study. Duration and Place of Study: Armed Forces Institute of Pathology, Rawalpindi from Pakistan, Apr to Sep 2020. Methodology: All samples received at the Microbiology Department during the study period were included. Various samples such as blood, pus, sputum, Endobronchial washing (EBW), non-directed bronchial lavage (NBL), tissue and urine were processed in the laboratory. Samples were inoculated on appropriate culture media, and bacteria were identified according to their colony morphology, Gram staining characteristics and biochemical tests. Antibiotic susceptibility testing was performed using the Kirby Bauer Disk Diffusion technique according to Clinical and Laboratory Standards Institute (CLSI) 2020 guidelines. However, for susceptibility to Colistin, the agar dilution method was performed. Results: In our study, fifty-five Carbapenem-resistant Pseudomonas aeruginosa were isolated. Out of these fifty-five isolates, thirteen (23.6%) were sensitive to Tazocin, seven (12.7%) were sensitive to Ceftazidime, fourteen (25.4%) were sensitive to Gentamicin, 7(12.7%) were sensitive to Cefepime, (12.7%) to Aztreonam, 10(18.2%) were sensitive to Amikacin, 7(12.7%) to Ciprofloxacin, 6(11%) to Levofloxacin and forty-two (76.4%) were sensitive to Colistin. Conclusion: The susceptibility of Carbapenem-resistant Pseudomonas aeruginosa to all the antibiotics tested in this study was low. Isolates displayed the highest susceptibility to Colistin. [ABSTRACT FROM AUTHOR]

Meningitis can present as a stroke mimic. We report a case of a 69 years old lady who presented to accident and emergency 2-hours after an episode of vomiting followed by sudden onset left sided weakness and drowsiness. Examination revealed left sided weakness with dysphasia. Initial plan was to decide if the patient was for thrombolysis. An urgent CT scan ruled out hemorrhage. On re-examination the patient was febrile and there was some degree of neck stiffness. Magnetic resonance imaging of brain was performed (DW1 no stroke) which ruled out acute ischemia. A lumber puncture was done eventually that showed low glucose, high leukocyte count, and gram staining revealed diplococci in chains. Clinicians should bear in mind the differential diagnosis of meningitis when there are risk factors and overlapping clinical symptoms. [ABSTRACT FROM AUTHOR]

Objective: To compare the frequency of E.faecalis in the saliva and root canals of teeth associated with apical periodontitis due to endodontic treatment failure. Study Design: Cross-sectional comparative. Place and Duration of Study: Samples were collected from Operative Dentistry department, AFID, while laboratory processing was done at AFIP, Rawalpindi. Duration of this study was one year. Patients and Method: Fifty patients, both males and females with failed endodontic treatment were selected. Saliva and root canal samples were collected from each patient, inoculated on MacKonkey agar plate and incubated at 35-370C for 48 hours. E.faecalis colonies were identified by colony morphology, Gram stain, catalase, bile asculin test, arabinose fermentation and growth in 6% NaCl nutrient broth. Results: The frequency of E. faecalis in saliva was 34% and 58% in root canal samples. Frequency of the presence of E.faecalis in root canals and saliva was found to be statistically different (p=0.000). Conclusion: The presence of E.faecalis in root canal was not associated with their presence in saliva. [ABSTRACT FROM AUTHOR]