Synonymous and non-synonymous polymorphisms in toll-like receptor 2 (TLR2) gene among complicated measles cases at a tertiary care hospital, Peshawar, Pakistan.

OBJECTIVES: To detect single nucleotide polymorphism in toll-like receptor 2 (TLR2) gene in complicated cases of measles, in order to understand the genetic basis of complex human immune responses against measles complications. METHODS: A total of 100 children consisted of 50 measles complicated cases while rest were gender matched disease-free individuals who served as controls for this study. Patient demographic data and clinical information were recorded on a separate pre-designed model form. All exonic regions of TLR2 gene of the patients and control samples were amplified through polymerase chain reaction. Various in-silico mutation verification tools like protein variation effect analyzer, MUPRO, sorting intolerant from tolerant, functional analysis through hidden Markov models, and polymorphism phenotyping v2 to study the effect of novel non-synonymous polymorphism on structure and function of TLR2 protein. RESULTS: Synonymous and novel non-synonymous polymorphisms were identified in measles complicated cases. Among these, rs1816702 was marked to 5 untranslated region section of TLR2 gene, while rs3804099 and rs3804100 were identified in the coding region. Novel non-synonymous polymorphisms were shown in the coding region of TLR2 gene. No significant association was established between the observed genetic polymorphisms and measles complications. However, rs3804100 increased the risk of lower respiratory tract infection. CONCLUSION: The overall impact of novel non-synonymous polymorphism of TLR2 protein structure and functions was neutral and tolerated.

Phenotypic and molecular characterization of virulence factors of extra-intestinal pathogenic Escherichia coli isolated from patients of Peshawar, Pakistan.

Extra-Intestinal Escherichia coli (ExPEC) are important cause of Urinary Tract Infections (UTIs) and systemic infections. The purpose of this study was to investigate numerous ExPEC bacterial isolates for phenotypic virulence characteristics including hemolytic activity and resistance pattern and to observe their association with genetic traits via Polymerase Chain Reaction (PCR). A total of 367 ExPEC isolates were collected from patients admitted in Khyber Teaching Hospital (KTH) Peshawar, Pakistan. Standard techniques were used for identification of isolates, determination of hemolytic potential and antimicrobial susceptibility testing. PCR was used for screening of virulence genes using specific primers. A total of 367 ExPEC isolates were characterized, among which 62.7, 24.3, 7.1 and 6% were isolated from urine, pus, sputum and wound specimens, respectively. Majority of the isolates (82.8%) were hemolysin positive. Multi drug resistance pattern was shown by 41% of the isolates and harbored at least one virulence gene (71.7%), of which sat was the most prevalent (64.3%). The highest resistance was found to cefotaxime (99.2%), ampicillin (97.5%) and aztreonem (89.6%). 15 different virulence genes combinations were observed in the current study. A total of 16 virotypes (15 of positive virulence genes and one of no virulence gene) were observed in the current study. The current investigation showed a high prevalence of sat and hlyA genes among ExPEC isolate, suggesting a role of these genes in the pathogenesis of ExPEC.

The Resurgence of Measles Infection and its Associated Complications in Early Childhood at a Tertiary Care Hospital in Peshawar, Pakistan.

Measles infection is of substantial interest to immunologists due to its paradoxical interaction with the immune system. After the acquisition of the measles infection, secondary infection plays a pivotal role in measles-related deaths. A cross-sectional study conducted between December 2018 and July 2019 is presented here. A total of one hundred children of both genders presented with measles complications were included following WHO criteria. Measles confirmation was done by quantitative determination of anti-measles antibodies (IgM) in patients' sera while patient-related demographic data, vaccination status, and other clinical information were obtained on a separate form. The number of female patients (52%) slightly exceeded the number of males (48%). 43% of patients' parents were illiterate, and half of the patients (50%) were from a poor background. The majority of children (76%) who presented with the complications did not receive a measles vaccine. 56% of children were breastfed while 58% received vitamin A supplements but developed complications. The elevated levels of anti-measles IgM were observed in 77% of cases. In both genders, the major complications were pneumonia, lower respiratory tract infection (LRTI), acute diarrhea, diarrhea and LRTI, pneumonia and diarrhea, otitis media and pneumonia, myocarditis and LRTI, and pneumothorax. The majority of the infected children (n = 48) under 12 months of age had associated complications. It has been observed that the measles virus strikes early age children in the northwestern region of Pakistan, which is an alarming situation and is associated with the aforementioned complications, especially in unvaccinated children. Anti-measles IgM is an important serological parameter for early diagnosis of measles infection.Measles infection is of substantial interest to immunologists due to its paradoxical interaction with the immune system. After the acquisition of the measles infection, secondary infection plays a pivotal role in measles-related deaths. A cross-sectional study conducted between December 2018 and July 2019 is presented here. A total of one hundred children of both genders presented with measles complications were included following WHO criteria. Measles confirmation was done by quantitative determination of anti-measles antibodies (IgM) in patients' sera while patient-related demographic data, vaccination status, and other clinical information were obtained on a separate form. The number of female patients (52%) slightly exceeded the number of males (48%). 43% of patients' parents were illiterate, and half of the patients (50%) were from a poor background. The majority of children (76%) who presented with the complications did not receive a measles vaccine. 56% of children were breastfed while 58% received vitamin A supplements but developed complications. The elevated levels of anti-measles IgM were observed in 77% of cases. In both genders, the major complications were pneumonia, lower respiratory tract infection (LRTI), acute diarrhea, diarrhea and LRTI, pneumonia and diarrhea, otitis media and pneumonia, myocarditis and LRTI, and pneumothorax. The majority of the infected children (n = 48) under 12 months of age had associated complications. It has been observed that the measles virus strikes early age children in the northwestern region of Pakistan, which is an alarming situation and is associated with the aforementioned complications, especially in unvaccinated children. Anti-measles IgM is an important serological parameter for early diagnosis of measles infection.

Evaluation of GeneXpert MTB/RIF Assay for Detection of Pulmonary Tuberculosis on Sputum Samples.

OBJECTIVE: To analyse the diagnostic performance of MTB/RIF assay for the diagnosis of pulmonary tuberculosis and detection of rifampicin resistance using sputum samples. STUDY DESIGN: Observational cross-sectional study. PLACE AND DURATION OF STUDY: Provincial TB Reference Laboratory (PTRL), Hayatabad Medical Complex, Peshawar, Pakistan, from January to October 2015. METHODOLOGY: A total of 268 participants were consecutively enrolled in the study after meeting the inclusion criteria. Their sputum samples were collected and processed by N-acetyl-L-cysteine-sodium hydroxide (NALC-NaOH) method and GeneXpert MTB/RIF assay. RESULTS: This study determined the overall sensitivity and specificity of MTB/RIF assay, it was 92.4% (86/93) and 97.1% (138/142), respectively. The sensitivity was 98.4% (60/61) in culture proven smear positive samples, whilst sensitivity in culture proven smear negative samples was 93.7% (30/32), using culture as reference standard. CONCLUSION: GeneXpert MTB/RIF assay could greatly improve early diagnosis of PTB in smear negative cases as well as multidrug resistant tuberculosis.