Invasive fungal disease in critically ill and Immunocompromised patients.

OBJECTIVE: To study the frequency and type of invasive fungal disease in critically ill and immunocompromised patients. Method: The prospective, cross-sectional, descriptive study was conducted at the Armed Forces Institute of Pathology, Rawalpindi, Pakistan, from January 2017 to December 2020, and comprised pathological samples from immunocompromised and critically ill patients for fungal culture. Data regarding demographics, comorbidities, results of direct microscopy and fungal culture was recorded. Data was analysed using SPSS 22. RESULTS: Of the 8285 patients' specimens, 4722(57%) belonged to males and 3563(43%) to females. The mean age of the patients was 48.32±5.42 years (range: 14-98 years). Out of total 8285, 3465(41.82%) were related to blood, 2640(32%) endobronchial washing, 837(10%) sputum, 623(7.5%) tissue, 332(4%) body fluids, 288(3.5%) bronchoalveoar lavage and 100(1.2%) cerebrospinal fluid. Aspergillus flavus (20.7%) and candida albicans (14.5%) were the two most commonly isolated fungal species. CONCLUSIONS: A high index of suspicion for invasive fungal disease should be maintained in immunocompromised and critically ill patients.

Frequency of Cryptosporidiosis in Children having Persistent Diarrhea.

OBJECTIVE: Globally childhood diarrheal diseases continue to be the second leading cause of death. Cryptosporidium spp are important intestinal parasites that cause diarrhea in humans and animals particularly in developing countries. This investigation was carried out to find out the frequency of cryptosporidiosis in children presenting with persistent diarrhea. METHODS: Two hundred stool samples were collected in this descriptive cross-sectional study conducted at Microbiology Department, Combined Military Hospital, Lahore Pakistan between the months of July to Dec 2014. Children aged five years to 12 years who presented with persistent diarrhea were included in the study. Stool specimens were processed using the modified acid-fast staining method, and microscopically examined for Cryptosporidium infection. RESULTS: The average age of study participants was 7.95 with a standard deviation of 2.21 years. Among the participants 66% were males whereas 34% were females. Twenty eight percent had presence of oocysts in stool samples. CONCLUSIONS: The frequency of Cryptosporidiosis among children with persistent diarrhea was 28%. This high frequency indicates that this population is uniquely susceptible to infection. It also highlights the need for education about hygiene, accurate diagnosis, and treatment of Cryptosporidiosis. There is also a need for additional studies regarding the occurrence of this pathogen.

Multiplex PCR for Rapid Diagnosis of Drug Resistant Mycobacterium Tuberculosis.

OBJECTIVE: To evaluate a multiplex PCR for rapid diagnosis of drug resistant mycobacterium tuberculosis (MTB) strain. STUDY DESIGN: Cross-sectional observational study. PLACE AND DURATION OF STUDY: Department of Microbiology, Armed Forces Institute of Pathology (AFIP), Rawalpindi, from January to September 2018. METHODOLOGY: Over a period of 8 months, a total of 84 cultured positive samples were included in the study using nonprobability sampling techniques. MTB isolates were phenotypically characterised using MGIT 960 system for antituberculosis agents including rifampicin (RIF), isoniazid (INH), ethambutol (EMB) and Streptomycin. The DNA was extracted using Gentra system DNA extraction kit. The multiplex PCR was optimised for genetic characterisation of MTB samples for rpo B (rifampicin), kat G (isoniazid) and emb B (ethambutol) gene. The gel electrophoresis was performed to observe comparative banding pattern of amplified gene products. RESULTS: For detecting drug resistance, the specificity and sensitivity of multiplex PCR in isolates was 100% and 100% for rifampicin, 100% and 71% for isoniazid, and 100% and 60% for ethambutol, respectively. When compared to phenotypically resistance results, the positive predictive value (PPV) was 100% each and the negative predictive value (NPV) was calculated to be 100%, 74% and 71% for RIF, INH and EMB, respectively. CONCLUSION: Multiplex PCR is a useful gadget for quick determination of drug-resistant TB in specimens, hence permitting an initial therapeutic approach. However, for accurate management of patients, phenotypic method should be used to confirm results.

Efficacy and safety of artemehter-lumefantrine in uncomplicated falciparum malaria in Liberia.

OBJECTIVE: To assess the in vivo efficacy and adverse effects of Artemether-lumefantrine combination in acute uncomplicated falciparum malaria. METHODS: A prospective, observational study was conducted at Department of Medicine and Pathology, Pakistan Medical level II hospital, Tubmanburg and Harper from March 2009 to September 2009. One hundred subjects with positive Plasmodium falciparum rings on malaria slide fulfilled the selection criteria and were included in the study. Mean, minimum and maximum values along with standard deviation of age, malarial parasite index, fever clearance time and parasite clearance time were calculated. A 28 day parasitological cure rate was determined. Frequency of various adverse events observed during this study were also noted. RESULTS: Of 100 subjects, 70 wereAfricans while remaining were Asians. Mean fever clearance time in Africans and Asians were 18.9 +/- 11.5 and 27.9 +/- 14.3 hours respectively. The mean parasite clearance time was almost similar in both races ranging from 28 to 31 hours. A 28 day parasitological cure rate was found to be 100%. About 10% of the subjects developed mild to moderate side effects including headache, vomiting, loss of sleep, vertigo and diarrhoea. There was no mortality during the study. CONCLUSION: Artemether Lumefantrine combination therapy may be used safely and effectively in the management of acute uncomplicated falciparum malaria.

CTX-M ESBL enzyme in Escherichia coli from urology patients in Rawalpindi, Pakistan.

OBJECTIVE: To detect CTX-M phenotype utilizing disc diffusion and MIC testing in Escherichia coli isolated from a tertiary care urology setting. METHODS: Fifty single, non duplicate ESBL producing isolates from a tertiary care urology hospital were evaluated for the presence of CTX-M phenotype. Initially all the urinary isolates were tested for ESBL production. The isolates were identified by using API 20E galleries and screened for ESBL production by combination disc methods. Representative 4 ESBL isolates were sent to Antibiotic Resistance Monitoring and Reference Laboratory (ARMRL), Health Protection Agency, Colindale, London, UK where those were further subjected to MIC testing by agar dilution and E-test strips. RESULTS: A total of 4 ESBL producing E. coli isolates were characterized to be CTX-M on phenotypic characterization. The overall yield of CTX-M phenotypes was 75%. CONCLUSION: The emergence of CTX-M from Pakistan is alarming; however, further studies are required to study the epidemiology and genetic characterization of CTX-M types of ESBLs.