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.

An eye emergency at high altitude on the climber: things not to miss.

Risk of eye emergencies at high altitude should be noted by Emergency Physicians or expedition doctors for high altitude missions. Early management and understanding the sequelae of the conditions will make a difference in management and decisions at high altitude.

2 Handfuls of Medication in 2 Hours Just to Let Him Talk.

Repeated dose of adrenaline in anaphylaxis is limited evidence in clinical setting. Hence, the usage is depending on physician best interest and knowledge. We reported a case of repeated doses of adrenalin was given through nebulizer and intravenous in anaphylaxis. We believed the unusual circumstances of this case was likely to be repeated on some readers' clinical practice and this mode of treatment is an adjunct to consider in such cases especially in Emergency Department (ED).

Hematocolpos mimicking acute urinary retention in Emergency Department.

Lower abdominal pain with lower urinary tracts symptoms in young girls presented to Emergency Department (ED) is urinary tract infection (UTI) until proven otherwise. We reported a case of classic UTI presentation with unusual finding of hematocolpos. We believed the unusual circumstance of this case is likely to be repeated in some other clinical practice and such differential diagnoses should be considered.