Performance of an automated multiplex immunofluorescence assay for detection of Chlamydia trachomatis immunoglobulin G.

Chlamydia serology is indicated to investigate etiology of miscarriage, infertility, pelvic inflammatory disease, and ectopic pregnancy. Here, we assessed the reliability of a new automated-multiplex immunofluorescence assay (InoDiag test) to detect specific anti-C. trachomatis immunoglobulin G. Considering immunofluorescence assay (IF) as gold standard, InoDiag tests exhibited similar sensitivities (65.5%) but better specificities (95.1%-98%) than enzyme-linked immunosorbent assays (ELISAs). InoDiag tests demonstrated similar or lower cross-reactivity rates when compared to ELISA or IF.

Transnational authority in the knowledge-based economy: Who sets the standards of ICT training and certification?

This article examines the extent and limits of nonstate forms of authority in international relations. It analyzes how the information and communication technology (ICT) infrastructure for the tradability of services in a global knowledge-based economy relies on informal regulatory practices for the adjustment of ICT-related skills. By focusing on the challenge that highly volatile and short-lived cycles of demands for this type of knowledge pose for ensuring the right qualification of the labor force, the article explores how companies and associations provide training and certification programs as part of a growing market for educational services setting their own standards. The existing literature on non-conventional forms of authority in the global political economy has emphasized that the consent of actors, subject to informal rules and some form of state support, remains crucial for the effectiveness of those new forms of power. However, analyses based on a limited sample of actors tend toward a narrow understanding of the issues concerned and fail to fully explore the differentiated space in which non state authority is emerging. This article develops a three-dimensional analytical framework that brings together the scope of the issues involved, the range of nonstate actors concerned, and the spatial scope of their authority. The empirical findings highlight the limits of these new forms of nonstate authority and shed light on the role of the state and international governmental organizations in this new context.

Discordant prevalence of hypertension using two different automated blood pressure measurement devices: a population-based study in Dar es Salaam (Tanzania).

OBJECTIVE: The estimation of blood pressure is dependent on the accuracy of the measurement devices. We compared blood pressure readings obtained with an automated oscillometric arm-cuff device and with an automated oscillometric wrist-cuff device and then assessed the prevalence of defined blood pressure categories. METHODS: Within a population-based survey in Dar es Salaam (Tanzania), we selected all participants with a blood pressure >/= 160/95 mmHg (n=653) and a random sample of participants with blood pressure <160/95 mmHg (n=662), based on the first blood pressure reading. Blood pressure was reassessed 2 years later for 464 and 410 of the participants, respectively. In these 874 subjects, we compared the prevalence of blood pressure categories as estimated with each device. RESULTS: Overall, the wrist device gave higher blood pressure readings than the arm device (difference in systolic/diastolic blood pressure: 6.3 +/- 17.3/3.7 +/- 11.8 mmHg, P<0.001). However, the arm device tended to give lower readings than the wrist device for high blood pressure values. The prevalence of blood pressure categories differed substantially depending on which device was used, 29% and 14% for blood pressure <120/80 mmHg (arm device versus wrist device, respectively), 30% and 33% for blood pressure 120-139/80-89 mmHg, 17% and 26% for blood pressure 140-159/90-99 mmHg, 12% and 13% for blood pressure 160-179/100-109 mmHg and 13% and 14% for blood pressure >/= 180/110 mmHg. CONCLUSIONS: A large discrepancy in the estimated prevalence of blood pressure categories was observed using two different automatic measurement devices. This emphasizes that prevalence estimates based on automatic devices should be considered with caution.

Insertion of green fluorescent protein into nonstructural protein 5A allows direct visualization of functional hepatitis C virus replication complexes.

Hepatitis C virus (HCV) replicates its genome in a membrane-associated replication complex, composed of viral proteins, replicating RNA and altered cellular membranes. We describe here HCV replicons that allow the direct visualization of functional HCV replication complexes. Viable replicons selected from a library of Tn7-mediated random insertions in the coding sequence of nonstructural protein 5A (NS5A) allowed the identification of two sites near the NS5A C terminus that tolerated insertion of heterologous sequences. Replicons encoding green fluorescent protein (GFP) at these locations were only moderately impaired for HCV RNA replication. Expression of the NS5A-GFP fusion protein could be demonstrated by immunoblot, indicating that the GFP was retained during RNA replication and did not interfere with HCV polyprotein processing. More importantly, expression levels were robust enough to allow direct visualization of the fusion protein by fluorescence microscopy. NS5A-GFP appeared as brightly fluorescing dot-like structures in the cytoplasm. By confocal laser scanning microscopy, NS5A-GFP colocalized with other HCV nonstructural proteins and nascent viral RNA, indicating that the dot-like structures, identified as membranous webs by electron microscopy, represent functional HCV replication complexes. These findings reveal an unexpected flexibility of the C-terminal domain of NS5A and provide tools for studying the formation and turnover of HCV replication complexes in living cells.

Automated DNA extraction using the QIAsymphony platform: Estimation of DNA recovery from simulated forensic stains

The aim of this study was to evaluate the forensic protocol recently developed by Qiagen for the QIAsymphony automated DNA extraction platform. Samples containing low amounts of DNA were specifically considered, since they represent the majority of samples processed in our laboratory. The analysis of simulated blood and saliva traces showed that the highest DNA yields were obtained with the maximal elution volume available for the forensic protocol, that is 200 ml. Resulting DNA extracts were too diluted for successful DNA profiling and required a concentration. This additional step is time consuming and potentially increases inversion and contamination risks. The 200 ml DNA extracts were concentrated to 25 ml, and the DNA recovery estimated with real-time PCR as well as with the percentage of SGM Plus alleles detected. Results using our manual protocol, based on the QIAamp DNA mini kit, and the automated protocol were comparable. Further tests will be conducted to determine more precisely DNA recovery, contamination risk and PCR inhibitors removal, once a definitive procedure, allowing the concentration of DNA extracts from low yield samples, will be available for the QIAsymphony.

Angiofil-mediated visualization of the vascular system by microcomputed tomography: a feasibility study.

Visualization of the vascular systems of organs or of small animals is important for an assessment of basic physiological conditions, especially in studies that involve genetically manipulated mice. For a detailed morphological analysis of the vascular tree, it is necessary to demonstrate the system in its entirety. In this study, we present a new lipophilic contrast agent, Angiofil, for performing postmortem microangiography by using microcomputed tomography. The new contrast agent was tested in 10 wild-type mice. Imaging of the vascular system revealed vessels down to the caliber of capillaries, and the digital three-dimensional data obtained from the scans allowed for virtual cutting, amplification, and scaling without destroying the sample. By use of computer software, parameters such as vessel length and caliber could be quantified and remapped by color coding onto the surface of the vascular system. The liquid Angiofil is easy to handle and highly radio-opaque. Because of its lipophilic abilities, it is retained intravascularly, hence it facilitates virtual vessel segmentation, and yields an enduring signal which is advantageous during repetitive investigations, or if samples need to be transported from the site of preparation to the place of actual analysis, respectively. These characteristics make Angiofil a promising novel contrast agent; when combined with microcomputed tomography, it has the potential to turn into a powerful method for rapid vascular phenotyping.