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: Dermatophytoses infections are widespread in the developing world. The laboratory diagnosis of dermatophytes has been a challenge as it involves microscopy and trained personnel. Potassium hydroxide wet mount with dimethyl sulfoxide added is routinely used in direct microscopy. But it lacks color contrast and the hyphae may be missed on routine microscopy. The study aimed to evaluate the effectiveness of the Chicago sky blue stain against routine potassium hydroxide-dimethyl sulfoxide (KOH/DMSO) wet mount in direct microscopy. Patients and methods: The study was carried out at the Department of Microbiology, Postgraduate Medical Institute, Lahore over a period of nine months from July 2013 till March 2014. Patients of either gender regardless of age, clinically diagnosed as having dermatophytoses by dermatologists were selected for this study. Specimens from 100 patients were collected from the dermatology outdoor of a tertiary care hospital for this study. They were evaluated microscopically with routine potassium hydroxide-dimethyl sulfoxide (KOH-DMSO) wet mount and Chicago sky blue (CSB) stain. Data were collected and entered in Statistical Package for the Social Sciences (SPSS) version 20.0. Results: Out of a total of 100 samples collected from skin, hair and nails, 59% were positive on direct microscopy with KOH/DMSO wet mount. Whereas direct microscopy using CSB stain revealed dermatophytes in 62% of cases. Conclusion: Chicago sky blue staining is a better technique for the detection of dermatophytes as compared to potassium hydroxide wet mount examination. It is simple, rapid, and easy to interpret. We recommend the use of this technique to improve the detection of dermatophytes without awaiting the results of the culture.
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.
2 μg/ml were screened for heteroresistance by Glycopeptide Resistance Detection (GRD) E-test and Vancomycin screen agar. Data was entered and analyzed by using SPSS version 20.0. Results: When compared with E test GRD, Vancomycin screen agar (V3) showed 100% sensitivity with a 95% CI 39.76% to 100% and the specificity was 65 % with a 95 % CI 47.46% to 79.79%. Its PPV was 23% and NPV was 100% with an overall diagnostic accuracy of 68%. When compared with E test GRD, Vancomycin screen agar (V4) showed a sensitivity of 75% with a 95% CI 19.41% to 99.37% and a specificity of 86.47% with a 95% CI 71.91 to 95.59%. Its PPV was 37.5% and NPV predictive value was 96.96% with an overall diagnostic accuracy of 85.36%. Conclusion: In developing countries like Pakistan, where E tests are costly and difficult to use in routine laboratories, a screening test, which does not miss heteroresistant VISA may be of clinical use.
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