Development and validation of TaqMan real-time PCR for the detection of Burkholderia pseudomallei isolates from Malaysia

@#Rapid detection of Burkholderia pseudomallei, the etiologic agent of melioidosis, allows for timely initiation of appropriate treatment and better clinical outcomes. In the current gold standard, the culture method is time consuming and suffers from low sensitivity. Meanwhile, previously reported molecular assays are fast and sensitive, but their performance on isolates from Malaysia, an endemic region of melioidosis is under reported. This study designed oligonucleotides targeting orf2 of Type III secretion system (TTSS) genes cluster for the detection of Malaysian B. pseudomallei isolates and evaluated the assay on 95 local B. pseudomallei strains, 58 other microorganisms and 71 clinical specimens from patients. The developed assay exclusively detected all tested B. pseudomallei isolates with a detection limit of 20 fg per reaction (equivalent to ~2.5 copies). Subsequent testing on clinical samples showed that the assay detected all confirmed specimens with the growth of B. pseudomallei (n = 10/10). None of the negative specimens had a detectable signal of our TTSS-orf2 assay (n = 0/61). In conclusion, the present study provides crucial preliminary data for a subsequent study and should be considered as a potential alternative to current time-consuming culture method for the detection of B. pseudomallei.

Aedes albopictus in urban and forested areas of Malaysia: A study of mitochondrial sequence variation using the CO1 marker

@#This study explores the use of a long fragment of the mitochondrial cytochrome oxidase subunit 1 (CO1) marker to elucidate the genetic diversity of Aedes albopictus sampled from urban and forested regions in Peninsular and East Malaysia. A total of 36 samples were collected from 5 localities from which its genetic variability was analysed. 33 distinct mtDNA haplotypes were identified following the amplification and sequencing of the concatenated CO1 gene. The analysed region of the CO1 gene identified substantial levels of genetic diversity among mosquitoes in urban populations and revealed unique genealogical relationships between local isolates as revealed in the haplotype network. This study highlights the reliability of the long CO1 fragment to identify genetic divergence of Aedes albopictus which can be utilized in forthcoming studies.

Study of growth in prepubertal asthmatics.

OBJECTIVE: The aim of this pilot study was to assess whether long standing asthma affects growth in prepubertal Egyptian children before initiation of long-term corticosteroid therapy. METHODS: Children with asthma were divided into two groups according to asthma severity, moderate (n=24) and severe (n=14) and were compared for their physical and skeletal growth with a control group (n=15) using standard deviation score (SDS) and one-way ANOVA (analysis of variance) test. RESULTS: No statistically significant differences were found between various growth parameters (weight, height, BMI, upper segment lower segment ratio, and skin fold thickness in asthmatic and normal children, although a positive correlation was found between the age at which the asthma presented and the height in all asthmatic children, r= 0.288, p= 0.036. The bone age standard deviation scores (SDS) were 0.97 mean, -0.165 and -0.572 for controls, moderate and severe asthmatics respectively (P< 0.05), and significant inter group difference between the 2 asthmatic groups (moderate and severe) and the controls was found. CONCLUSION: The authors conclude that there was no significant major effect of asthma per se on growth parameters in children, but that skeletal maturation was influenced by long standing asthma.

Evaluation of the efficacy, safety and tolerability of valdecoxib gel (1%) in adult patients with painful inflammatory joint disease.

Valdecoxib, a COX-2 inhibitor, has recently been introduced as a gel formulation. The present study was conducted to evaluate the efficacy, safety and tolerability of valdecoxib gel in adult patients with painful inflammatory joint conditions. The present study was a 10-day prospective, open, multicentric (6 centres) trial. Patients with clinical and radiological diagnosis of painful inflammatory joint conditions were treated with valdecoxib gel (1%). Efficacy was assessed by visual analogue scale (VAS), patient's and physician's global assessment of pain relief. Grading of associated clinical manifestations such as stiffness, swelling, tenderness and restriction of mobility was done. Tolerability and safety was assessed by physical examination, laboratory parameters and evaluation of adverse events. There was a statistically significant decrease in the mean pain visual analogue score (p<0.05). Onset of pain relief was within 15 minutes. There was a reduction of 58.8%, 57.2%, 65.4% and 60.2% in mean scores of stiffness, swelling, tenderness and mobility respectively from the baseline which was statistically significant. The laboratory values were within normal limits. The drug was well tolerated. There was no report of any hypersensitivity reaction. This study confirms that valdecoxib gel (1%) is an effective and safe option for the management of painful inflammatory joint conditions.