BackgroundVulvovaginal candidiasis (VVC) is considered as a pervasive gynecological problem among women worldwide. Owing to this fact, in the current study, we aimed at assessing the prevalence rate of Candida spp. causing VVC in symptomatic pregnant women and their antifungal susceptibility pattern.MethodsThis study was carried out in the tertiary care hospitals of Peshawar during the period of July 1, 2016 to December 31, 2016. The study group included 450 pregnant women in the age group of 17–44 years with symptoms of excessive vaginal discharge, pain and pruritis. In all, 108 pregnant women were culture positive for Candida. Antimicrobial susceptibility testing (AST) was conducted on specimens against various azoles and polyene F group of antifungals.ResultsOut of 108 Candida spp. isolated from vaginal swabs, there were 45 (41.7%) Candida albicans, 18 (16.7%) Candida tropicalis, 18 (16.7%) Candida krusei, 16 (14.8%) Candida glabrata and 11 (10.2%) Candida dubliniensis. According to age distribution, 27 years was the mean age. Pregnancy trimester distribution among patients was as follows: 21 (19.4%) patients were in their first trimester, 65 (60.2%) patients were in their second trimester and 22 (20.4%) patients were in the third trimester. Susceptibility of fluconazole was determined as follows: 33.3% of the Candida isolates were sensitive, 4.6% were susceptible dose dependent (SDD) and 62% were resistant. Susceptibility of Candida spp. with respect to nystatin in patients with VVC was as follows: 25% were sensitive, 16.7% were SDD and 58.3% were resistant. Susceptibility of clotrimazole was analyzed, and it was sensitive in 21.3% of patients, SDD in 19.4% of patients and resistant in 59.3% of patients. Voriconazole susceptibility was recorded to be sensitive in 85.2% of patients, SDD in 4.6% of patients and resistant in 10.2% of patients suffering from VVC. Susceptibility results for itraconazole in patients with VVC were as follows: 42.6% of patients were sensitive, 16.7% of patients were SDD, and 40.7% of patients were resistant.ConclusionIn this study, frequency of VVC was noted to be high in the second trimester of pregnancy, with the highest frequency of C. albicans isolated, followed by C. tropicalis and C. krusei. Antifungal susceptibility testing revealed that fluconazole was exceedingly resistant against Candida species (62%), followed by clotrimazole (59.3%) and nystatin (58.3%). On the contrary, voriconazole had the highest antimicrobial activity against Candida species (85.2%).
BackgroundThe purpose of this study was to explore molecular epidemiology of HCV genotype 3a in Peshawar based on sequencing and phylogenetic analysis of Core region of HCV genome.MethodsChronically infected Hepatitis C virus infected patients enrolled under the Prime Minister Hepatitis C control program at three Tertiary care units of Peshawar [Khyber Teaching Hospital Peshawar, Lady Reading Hospital Peshawar, Hayat Abad Medical Complex Peshawar] were included in this cross sectional observational study. Qualitative detection of HCV and HCV genotyping was carried out by a modified reverse transcription-polymerase chain reaction (RT-PCR) and type specific genotyping assay. The Core gene of HCV genotype 3a was amplified, cloned and sequenced. The sequences obtained were used for phylogenetic analysis using MEGA 6 software.ResultsAmong the 422 (82.75 %) PCR positive samples, 192 (45.5 %) were identified as having HCV genotype 3a infection. HCV Core gene sequencing was carried out randomly for the characterization of HCV 3a. Nucleotide sequence analysis of the obtained viral genomic sequences based on partial HCV 3a Core gene sequences with reference sequences from different countries showed that our sequences clustered with some local and regional sequences with high bootstrap values.ConclusionHCV 3a is highly prevalent in Peshawar, Pakistan and its phylogenetics based on Core gene sequences indicate the prevalence of different lineages of HCV 3a in Peshawar which may have consequences for disease management strategies causing more economic pressure on the impoverished population due to possible antiviral resistance.
The drug resistance genes are responsible to preserve the Pseudomonas. This also happens in the case of Mex drug efflux pumps and expression increase of MexA, MexB and OprM genes in Pseudomonas aeruginosa, when grown in sub-inhibitory concentrations of antibiotics. Objectives: This study was designed to detect the MexA gene of Pseudomonas aeruginosa resistant strains in tertiary care hospital, Peshawar. Study Design: Cross-sectional study. Setting: Department of Pathology, Khyber Teaching Hospital, Peshawar, Pakistan. Period: 14 months duration from April 2015 to May 2016. Material & Methods: The specimens including burn wound swabs, pus and urine were obtained from different patients and were processed on blood agar and MacConkey medium for isolation and identification. Conventional PCR was performed for the MexA gene on 50 specimens. Results: The simple conventional PCR was done for MexA (The Mex AB OprM operon resistance genes) and the O-antigen acetylase gene (the species-specific gene) separately, gave positive bands for 49 out of the 50 specimens. Our finding confirms the presence of the MexA gene (and hence most probably MexABOprM operon) in 49 out of 50 specimens of Pseudomonas aeruginosa. Conclusion: Among other resistance mechanisms to antibiotics and disinfectants, the MexABOprM efflux pump might have a role.
Background: Hepatitis C virus (HCV) belongs to the genus Hepacivirus and family Flaviviridae and is a leading cause of chronic liver diseases including cirrhosis and hepatocellular carcinoma. Objectives: This study aimed to analyze the genetic diversity of HCV genotype 3a based on complete core protein in Peshawar. Methods: Hepatitis C virus genotype 3a infected patients, belonging to different areas of Peshawar participated in this study. The complete core gene of HCV 3a was amplified and sequenced. The obtained sequences were used for mutational and phylogenetic analysis using CLUSTAL W and MEGA 6 software. Results: Phylogenetic analysis revealed that most of the HCV 3a strains prevalent in Peshawar are genetically closer to HCV 3a strains previously reported from Pakistan and India. Analysis of translated amino acids aligned against reference 3a strain (NZL1) demonstrated substitutions in three functional domains (D1, D2, D3) of the core protein. Core protein mutations R70Q (arginine to glutamine) and L/C91M (leucine/cysteine to methionine) were not found among studied isolates. In sequence PK/59 at position 72, glutamic acid was replaced with aspartic acid. Position 182 was occupied by leucine in place of phenylalanine in all sequences. Alanine and serine amino acids at the positions of 189 and 191 were replaced with threonine and cysteine, respectively. In seven of our isolates (PK/59-PK/68) glutamic acid and serine were found mutated to aspartic acid and cysteine, respectively. Conclusions: HCV core protein, although a conserved region could be considered a phylogenetic marker for the determination of genetic relatedness among circulating viral strains and tracing the outbreak of an infection. Geographical, regional, and genotypic differences are effective in the prevalence of substitutions in HCV core protein.
Background: Cystic echinococcosis is a zoonotic, neglected tropical disease caused by various genotypes of cestode parasite Echinococcus granulosus. Although, echinococcosis causes enormous financial and health impacts, no gold standard diagnostic technique is available. Genetic characterization of these prevalent cestode at species level is essential for disease management and appropriate control measures. Objective: We aimed to investigate the genetic diversity of echinococcus granulosus using a modified Polymerase Chain Reaction-Restriction Fragment Length Polymorphism (PCR-RFLP) based assay in infected population. Methodology: A total of 18 human hydatid cyst samples were collected from various hospitals of Southern Punjab and Islamabad Capital Territory region of Pakistan. Extracted DNA was used for PCR amplification of mitochondrial NADH dehydrogenase subunit 1 (Nad1) gene followed by sequencing and phylogenetic analysis using Molecular evolutionary Genetic Analysis (MEGA) Software. The entire sequences were fed into NEBcutter V2.0 to select a single restriction enzyme followed by invitro confirmation through PCR-RFLP. Results: Amplification on the Nad1 gene was observed in 100% of the samples processed. The Basic Local Alignment Tool (BLAST) and phylogenetic tree analysis revealed 83.3% E. granulosus (G1-G3 genotypes), 11.1% E. multilocularis and 5.6% E. Canadensis (G6 genotype). The use of the BfaI enzyme in PCR-RFLP analysis revealed that all of the 18 samples were assigned consecutive genotypes as observed in the sequencing. Conclusion: The current study concluded that the BfaI enzyme could be used for the genotypic analysis of echinococcosis in developing and frequently affected countries. It will be a cost-effective and easy technique compared to sequencing, which will aid in developing novel therapeutic and control strategies for the parasite.
Toxoplasmosis and Brucellosis are zoonotic infections having worldwide distribution. Toxoplasma gondii and Brucella species are the causative agents of these infections. The human can be infected by contact with infected animals. These infections affect the reproductive system of humans and animals. The study aimed to determine the prevalence of Toxoplasma gondii and Brucella species in infertile women of District Dir Lower. A cross-sectional study was conducted in General Hospital and Maternity Home District Dir Lower and a total of 576 infertile women were screened for the detection of Toxoplasma gondii and Brucella antibodies. ICT method was used for the detection of Anti-T. gondii and the detection of Brucella antibodies, the serum agglutination method was used. The overall prevalence of T.gondii was 11.3% (IgG=10.6% and IgM=0.7%), Brucella Abortus was 14.5% and the prevalence of Brucella Melitensis was 15.1%. The highest prevalence of T.gondii was observed among the age group 26-34 years while the lowest prevalence was observed among the age group 15-25 years. The highest prevalence of Brucella species was observed among the age group 35-45 years and the lowest prevalence was observed among the age group 15-25 years, and the prevalence of Brucella species was higher than T. gondii.
Hepatitis C Virus (HCV) is a flavivirus responsible for causing chronic liver diseases including cirrhosis and hepatocellular carcinoma. Identification of various patient and virus-related factors that can help predict response to antiviral therapy is extremely important in formulating the best therapeutic strategy for each patient either to continue or stop the therapy. The present study aimed to determine Sustained Virological Response (SVR) in HCV genotype 3a infected patients that received combination therapy of Sofosbuvir and Ribavirin and to investigate various factors that can help predict SVR. Study Design: Longitudinal Study Settings: Institute of basic Medical Sciences, Khyber Medical University, Peshawar (IBMS, KMU). Period: July 2016 to September 2017. Materials and Methods: Treatment response was evaluated among 100 HCV genotype 3a infected patients that received Sofosbuvir and Ribavirin therapy for 24 weeks. Various baseline parameters including hematological, biochemical and viral profiles of were recorded. HCV genotype determination was carried out by type specific nested PCR based genotyping assay. Viral load was determined at baseline, at 12 weeks for Early Virological Response (EVR) and at 24 weeks of treatment for SVR. Viral RNA quantification was carried out by Real Time PCR. Results: Out of 100 patients, SVR was observed in 83% of patients; while 17% of the chronic HCV 3a infected patients were Non-Responders (NR). Mean age of patients was low 34 ± 9.8 among patients who achieved SVR as compared to patients with non-response (41 ± 10). The 24 weeks Alanine aminotransferase (ALT) levels were significantly lower among patients with SVR (p-value ≤ 0.05). Although statistically not significant, baseline viral load was higher in NR group (p-value ≥ 0.05), than those with SVR. Association of EVR with SVR was found statistically significant (OR= 2.8, 95% CI 1.2-6.4, p-value ≤ 0.05). Conclusions: The current study indicated that pre and on-treatment monitoring of patients receiving anti-viral therapy is important for the management of patients with chronic HCV infection.
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