Toxoplasmosis

Western blotting is a valuable tool to studies ranging from regulatory signaling processes to confirmatory serum diagnosis of HIV [68 (link)–70 (link)]. The evolution of western blot technique from identification of a specific protein in a complex mixture to the direct detection of protein in a single cell allows this technique to be an important analytical tool for clinical research. An advanced single cell western blotting technique was employed to study stem cell signaling and differentiation as well as drug response in tumor cells [69 (link)]. Through single cell western blotting it was possible to analyze cell-to-cell variations in approximately 2000 cells simultaneously within complex populations of cells [71 (link)]. With the integration of intact cell imaging, the technique allows the identification of protein expression changes of a single drug resistant tumor cell and its isoforms among heterogeneous population of tumor cells in human glioblastoma cells treated with chemotherapeutic daunomycin [69 (link)]. Identification of upregulated multidrug resistant protein, P-glycoprotein in living glioblastoma subpopulations was indicative of an active drug eflux pump as an underlying mechanism for drug resistance [69 (link),71 (link)]. With the application of 2-DE gel separation together with spotting of protein by peptide mass fingerprint, the analysis of clinically relevant Helicobacter pylori (H. pylori) in related gastric disease conditions (chronic gastritis, duodenal ulcer) was possible [72 (link)]. The database of H. pylori (low expressed and membrane proteins) was created through the application of one-dimensional or 2-DE/MALDI-mass spectrometry techniques [72 (link)]. In a similar manner, the Simple Western technique was employed for the analysis of 15-valent pneumococcal vaccine PCV15-CRM197 [73 (link)]. Due to its high sensitivity and automation, the Simple Western method may be extended to analyze serotypes of other polysaccharide protein conjugate vaccines [73 (link)].
Western blotting is commonly used for the clinical diagnosis of various parasitic and fungal diseases including echinococcosis [74 (link)], toxoplasmosis [75 (link)], and aspergillosis [76 (link)]. In a recent study, the assay was successfully used for the reliable serodiagnosis of Farmer’s lung disease (FLD), a pulmonary disorder caused by inhalation of antigenic particles [77 (link)]. Thus, this technique can be exploited for rapid routine diagnosis of FLD in clinics [77 (link)]. Similarly, for immunodiagnostic of tuberculosis meningitis which is a chronic disease of central nervous system different molecular and immunological methods were used for clinical diagnosis of the disease. However, each of these techniques has their own limitations [78 (link)]. To overcome diagnostic issues of lower sensitivity and specificity, the immunoreactivity to Mycobacterium tuberculosis antigens was performed by western blotting [78 (link)]. Furthermore, western blotting was performed for the early and sensitive diagnosis of congenital toxoplasmosis [79 (link)] and was employed for rapid and sensitive serological diagnosis of a serious infectious disease paracoccidioidomycosis (PCM) [80 (link)]. Using immunoblotting, a new subgroup of human lymphotropic retroviruses (HTLV), was detected in patients with the acquired immunodeficiency syndrome (AIDS) [81 (link)]. Antigens of HTLV-III, specifically detected by antibodies in serum from AIDS or pre-AIDS patients [81 (link)]. Western blotting has also been used as a test for variant Creutzfeldt-Jakob Disease [82 (link)], some forms of Lyme disease [83 (link)] and is sometimes used as a confirmatory test for Hepatitis B [84 ] and Herpes Type 2 [85 (link)] infections. Western blots have also been used to confirm feline immunodeficiency status in cats [86 (link)].
Recently, a commercial Aspergillus western blotting IgG kit was developed by LD Bio Diagnostics (France) to carry out immunoblotting for the clinical diagnosis of chronic aspergillosis. The commercial kit was found to be sensitive and can analyze hundreds of samples from patients with aspergillus disease [87 (link)]. Thus, the clinical applications of western blotting technique will continue to progress as further advancements are made to improve sensitivity and reproducibility of the western blot.

The Cost-Effectiveness of Preventing AIDS Complications (CEPAC) International model is a state-transition model of HIV disease in resource-limited settings, with data derived for several country-specific analyses, including South Africa (16 (link), 20 (link), 21 (link)). Briefly, a cohort of hypothetical patients pass one at a time through “health states,” in monthly cycles, from entry into HIV care until death. Health states are defined to be both clinically and economically relevant and are stratified by current CD4 count, current HIV RNA level, and history of opportunistic disease. Opportunistic diseases are categorized into the following groups based on etiology, severity, and similarities in prophylaxis and treatment: mild or severe bacterial infections, mild or severe fungal infections, tuberculosis, toxoplasmosis, non-tuberculous mycobacteriosis, Pneumocystis jiroveci pneumonia, and other mild and severe diseases (5 (link)). Deaths in the model occur from acute opportunistic events (within 30 days of the event), chronic AIDS (not within 30 days of an opportunistic disease), or non-HIV-related causes (22 ).
Effective ART in the model functions to suppress HIV RNA and increase CD4 counts (23 (link), 24 (link)). Above and beyond the beneficial effect of increased CD4 count on opportunistic diseases and chronic HIV-related death (5 (link)), antiretroviral therapy per se results in an additional reduction in opportunistic diseases and chronic HIV-related death, as recently reported in Côte d'Ivoire and in the US (25 (link), 26 (link)). Clinical assessments are assumed to occur every 3 months, and CD4 and HIV RNA testing every 6 months while on therapy, consistent with South African recommendations (27 ). According to current standard of care, the model utilizes two sequential lines of antiretroviral therapy; the second-line is initiated when observed CD4 count decreases by 30% from its peak observed on-treatment level, or when a severe opportunistic disease is observed at least 6 months after initiating therapy (27 ). In accordance with current treatment guidelines, the second regimen for each patient is continued until death (7 , 28 (link)).