Molecular analysis of cross-bacterial contamination detected in biotin-free buffers during diagnosis of HCVinfections

In a routine work to determine Hepatitis C Virus (HCV) molecules in human infected serum with biotin/streptavidine enzyme linked immunosorbant assay (ELISA) technique, unexpected false positive was observed. No positive signals were noticed after changing all ELISA buffers. Subsequently, contaminated buffers were screened and analyzed for microbial contamination. Out of fifty-five, five randomly selected bacterial colonies were examined for biosynthesis of biotin using ELISA and/or Western blot binding biotin techniques in both supernatants and cell pellets. In compare to the E.coli reference strain a strong biotin signals in all examined isolates were recorded. All isolates were then examined for their genetic heterogeneity by PCR-RFLP technique of 23S rDNA genes. While, isolate BP(R2) which gave the highest biotin signal, was subjected for molecular identification. Comparative sequence analysis of the 16S rDNA gene (~1440 bp) revealed that this isolate is a member of the bacterial genus Delftia exhibiting a similarity value of 99.3% with Delftia acidovorans . In conclusion, a soluble biotin is secreted by the isolate Delftia acidovorans BP(R2) and it is also coupled to protein with molecular weight 25-26 KDa.As well as, this bacterial contaminationwas the reason for the false positive results observed during the detection of HCV infections. @JASE

Role of human resources management practices in the localisation of nursing workforce in Oman

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Role of human resources management practices in the localisation of nursing workforce in Oman

Omanisation is a national programme that focuses on creating job opportunities for the citizens of Oman by training and employing Omani nationals in jobs previously held by expatriates. In this study the aim was to explore the role of human resources management (HRM) practices in enhancing the success of nursing Omanisation. A mixed methodology approach was used to obtain the data needed for this study, starting with an extensive review of the literature, followed by an empirical field work comprising of interviews with senior healthcare and nursing managers, as well as a survey of nursing staff of various Omani healthcare organisations. Nine HRM practices were initially selected purposely from the literature. The relevance of these practices to nursing Omanisation was then confirmed by senior healthcare managers interviewed. One further HRM practice was identified. Through the interviews and survey all of these ten HRM practices were found to be in use but with varied effectiveness. The faster the Omanisation pace, the more it limited the development and preparation of Omani nurses to effectively take over from their expatriate counterparts. Challenges in implementing nursing Omanisation and its HRM related practices were identified, these included: managing young and inexperienced graduates; shortage of qualified staff; centralised Omanisation and career systems; managing cultural-work conflicts; and Omanising speciality areas. Moreover, the survey of nurses showed negative perceptions, particularly in relation to their preparation to take over from their expatriate counterparts, as well as their promotion and career progression. In the light of these findings, this study has proposed several recommendations, including gradual implementation of nursing Omanisation; review of expatriates' employment contracts; more emphasis on speciality training; resolving staff shortages; and review of the promotion and career progression systems

Selecting an appropriate method for expressing S locus F-box-S2 recombinant protein

A single locus (S locus) including at least two linked genes (female and male determinants) genetically controls the gametophytic self-incompatibility (GSI) in apple, which has evolved to avoid self-fertilization. There has been extensive work done on the female determinant of self-incompatibility, which has led to the determination of the tertiary structure of S-RNase. However, the tertiary structure of male determinant (S locus F-box, SLF/SFB) remains unresolved, which could mainly be due to difficulties associated with its expression in the recombinant expression systems. In addressing this, we have evaluated several in vivo (prokaryotic and eukaryotic) and in vitro expression systems for their efficiency in the expression of apple SLF2. The most successful expression of SLF2 (1 mg/ml) was achieved in E. coli using the synthesized gene in a high salt culture and applying heat shock before induction of culture. We therefore present an approach for the efficient expression of S locus F-box recombinant proteins for future functional and structural studies

Molecular analysis of cross-bacterial contamination detected in biotin-free buffers during diagnosis of HCV infections

In a routine work to determine Hepatitis C Virus (HCV) molecules in human infected serum with biotin/streptavidine enzyme linked immunosorbant assay (ELISA) technique, unexpected false positive was observed. No positive signals were noticed after changing all ELISA buffers. Subsequently, contaminated buffers were screened and analyzed for microbial contamination. Out of fifty-five, five randomly selected bacterial colonies were examined for biosynthesis of biotin using ELISA and/or Western blot binding biotin techniques in both supernatants and cell pellets. In compare to the E.coli reference strain a strong biotin signals in all examined isolates were recorded. All isolates were then examined for their genetic heterogeneity by PCR-RFLP technique of 23S rDNA genes. While, isolate BP(R2) which gave the highest biotin signal, was subjected for molecular identification. Comparative sequence analysis of the 16S rDNA gene (~1440 bp) revealed that this isolate is a member of the bacterial genus Delftia exhibiting a similarity value of 99.3% with Delftia acidovorans . In conclusion, a soluble biotin is secreted by the isolate Delftia acidovorans BP(R2) and it is also coupled to protein with molecular weight 25-26 KDa.As well as, this bacterial contaminationwas the reason for the false positive results observed during the detection of HCV infections. @JASE

Molecular analysis of cross-bacterial contamination detected in biotin-free buffers during diagnosis of HCV infections

In a routine work to determine Hepatitis C Virus (HCV) molecules in human infected serum with biotin/streptavidine enzyme linked immunosorbant assay (ELISA) technique, unexpected false positive was observed. No positive signals were noticed after changing all ELISA buffers. Subsequently, contaminated buffers were screened and analyzed for microbial contamination. Out of fifty-five, five randomly selected bacterial colonies were examined for biosynthesis of biotin using ELISA and/or Western blot binding biotin techniques in both supernatants and cell pellets. In compare to the E.coli reference strain a strong biotin signals in all examined isolates were recorded. All isolates were then examined for their genetic heterogeneity by PCR-RFLP technique of 23S rDNA genes. While, isolate BP(R2) which gave the highest biotin signal, was subjected for molecular identification. Comparative sequence analysis of the 16S rDNA gene (~1440 bp) revealed that this isolate is a member of the bacterial genus Delftia exhibiting a similarity value of 99.3% with Delftia acidovorans. In conclusion, a soluble biotin is secreted by the isolate Delftia acidovorans BP(R2) and it is also coupled to protein with molecular weight 25-26 KDa. As well as, this bacterial contamination was the reason for the false positive results observed during the detection of HCV infections. Journal of Applied Sciences and Environmental Management Vol. 9(1) 2005: 5-1