ABSTRACT
The purpose of this study was to develop a real time polymerase chain reaction (PCR) assay for the detection of the JAK2 V617F mutation that could be used in diagnostic laboratories.Sanger sequencing and a newly developed locked nucleic-acid, real-time PCR assay were used to detect the JAK2V617F mutation. There was 100% agreement between the sequencing and PCR analysis. Both assays were able to detect the mutation in all 24 of the 60 test specimens harbouring the mutation
Subject(s)
Myeloproliferative Disorders , Nucleic Acids , Real-Time Polymerase Chain Reaction , South AfricaABSTRACT
Acinetobacter baumannii is an important cause of hospital-acquired infections. The occurrence of carbapenem resistance that is caused by the carbapenem-hydrolysing class D ?-lactamases and the metallo-?-lactamases (MBLs) limits the range of therapeutic alternatives in treating A. baumannii infections. In this study; two multiplex polymerase chain reactions were performed to screen for both carbapenem-hydrolysing class D ?-lactamases and MBL genes in 97 clinical isolates of A. baumannii. Oxacillinase (OXA)-51 had a prevalence of 83 (81/97); and OXA-23 had a prevalence of 59 (57/97). One isolate was positive for an MBL [Verona integron-encoded metallo ?-lactamases (VIM)]. Therefore; continuous surveillance and monitoring of A. baumannii is crucial because of the high prevalence of antibiotic resistance genes
Subject(s)
Acinetobacter baumannii , Carbapenems , Cross Infection , Drug Resistance , PrevalenceABSTRACT
The hypervirulent polymerase chain reaction (PCR) ribotype 027 strain of Clostridium difficile produces toxins A; B and a binary toxin. Toxin detection kits are commonly used in diagnostic laboratories; but have been unsuccessful in detecting all of the relevant C. difficile strains; and the toxins produced. In this study; conventional PCR was used to detect the presence of the genes of toxin A; toxin B and the binary toxin of C. difficile. Eighty-four frozen (collected between 2006-2007) and 13 fresh (collected in 2010) stool specimens; obtained in Pretoria; were analysed. The genes for toxin A; toxin B and the binary toxin were detected in one of the fresh stool specimens. This may have implications for healthcare facilities; and suggests the possible emergence of the highly virulent PCR ribotype 027 strain of C. difficile in Pretoria. This emphasises the importance of continuous surveillance and monitoring of C. difficile outbreaks