ABSTRACT
Objective To understand correctly the hook like effect of the immune transmission and transmission turbidity in the automatic biochemical analyzer and the prevention of the hook like effect .Methods First ,it had the function set of measurement linear range of the upper limit of the instrument and the instrument automatic dilution redo ,and then ,had determination of excee‐ding the upper limit of measurement range and artificial dilution before the sample setting .Results The group average value was 2 .49 g/Lbefore the determination setting ,3 .31 g/L after the determination setting ,and 3 .33 g/L artificial dilution determination . Conclusion High dose hook effect makes the strong positive to weak positive samples of measurement by error ,even false negative results ,and high concentration sample makes low value .So the set of samples and dilution to determination of instrument ,measured values are more close to the true value .
ABSTRACT
Se hizo una valoración del impacto de los ensayos inmunoenzimáticos en la analítica de base inmunoquímica en las últimas 4 décadas, en la detección de agentes infecciosos o los productos asociado a su presencia y(o) actividad patogénica. Además se hace una incursión en algunos diseños y formatos que han tenido estos inmunoensayos desde los métodos electroquímicos de detección, los ensayos para detectar actividad proteolítica de origen microbiano y sus inhibidores como posibles blancos terapéuticos, los inmunoensayos directos de triple anticuerpo para lograr mayor sensibilidad, reveladores alternativos de la actividad enzimática, ensayos para el estudio de la serología viral con un mínimo de determinaciones, así como ensayos de competencia para evaluar la efectividad de candidatos vacunales basados en combinaciones peptídicas seleccionadas. Se concluyó con una rápida visión del futuro inmediato de este tipo de inmunoensayos a la luz de las tecnologías analíticas emergentes de detección.
This paper assessed the impact of the immunoenzymatic assays on the field of the immunochemistry-based analytics for the last 40 years, and on the detection of infectious agents or the products related to their presence and/or pathogenic activity. It also addressed some designs and formats of these immunoassays from electrochemical methods of detection, assays to determine proteolytic microbial activity and their inhibitors as possible therapeutical targets, more sensitive direct triple antibody systems, alternative enzymatic activity detectors, assays for viral serology of minimal determinations to competitive assays for evaluation of vaccinal candidate effectiveness based on selected peptide combinations. Finally, it provided a rapid overview of the near future of this type of immunoassays in the light of the emerging detection analytical technologies.
Subject(s)
Humans , Immunoenzyme Techniques/methods , Infections/microbiologyABSTRACT
La reacción hemolítica inmediata inmunológica representa la principal causa de muerte por transfusión. La reacción antígeno-anticuerpo con la subsiguiente hemólisis de los eritrocitos, desencadena un síndrome de respuesta inflamatoria sistémica como consecuencia de la liberación de citocinas (factor de necrosis tumoral a, interleucina 1, interleucina 8, interleucina 6) y activación de sistemas enzimáticos de amplificación (coagulación, fibrinólisis, calicreína-bradicinina). La coagulación intravascular diseminada y el fallo renal agudo son los eventos fisiopatológicos que dominan el cuadro. Se han desarrollado protocolos pre-transfusionales que ofrecen múltiples peldaños de seguridad, cuyo cumplimiento descarta la ocurrencia de reacciones fatales. Sin embargo, aún están presentes y se señalan como sus causas fundamentales los errores de identificación, la falta de conocimiento y la negligencia.
The immediate immunological hemolytic reaction is the main cause of death associated with transfusion. The antigen-antibody reaction with the subsequent .hemolysis of erythrocytes triggers a systemic inflammatory response syndrome resulting from the release of cytokines (tumoral a necrosis factor, interleukin 1, interleukin 8, interleukin 6) and from the activation of enzymatic amplification systems (coagulation, fibrinolysis, calicrein-bradykinin). The disseminated intravascular coagulation and the acute renal failure are physiopathological events that stand out in the clinical picture. Pretransfusional protocols that provide multiple safety steps, and whose fulfiment discards the occurrence of fatal reactions, have been developed. However, the idenfication erros, the lack of knowledge and negligence are still present and are considered as its fundamental causes.
ABSTRACT
Diagnosis by isolation of the microorganism from cultures provides the strongest evidence of infection by Paracoccidioides brasiliensis (Pb). However, isolation is not always possible and serological tests such as double immunodiffusion (ID) must be often employed. We analyzed the reaction profile of 75 serum samples from paracoccidioidomycosis (PCM) patients, by using ID, against nine different antigenic preparations of Pb: somatic antigen (SoAg), produced from Pb113 and Pb339 strains; cell-free antigen (CFAg), produced from Pb113 strain both preparations were cultured in Fava-Netto's agar medium for 7 days at 36°C; metabolic antigen (MAg), produced from Pb113 and Pb339 strains cultured in liquid NGTA (neopeptone, glucose, thiamine and asparagine) medium for 20 days at 36°C; soluble components of the cell wall outer surface (SCCWOS), produced from Pb113 strain cultured in Fava-Neto's agar medium at 36°C for 5, 10, 15, and 20 days; and Pb113 Negroni and Pb113 NGTA antigens, cultured for 20 days at 36°C. Serum samples reactivity was 90% to AgSo and SCCWOS cultured for 5, 10, 15 and 20 days; 86.6% to CFAg; 83.3% to MAg; 80% to Pb113 NGTA antigen; and 76.6% to Pb113 Negroni antigen. Electrophoresis in 10%SDS-PAGE showed high complexity of the protein fractions of SCCWOS, Pb113 Negroni and Pb113 NGTA antigens, which presented molecular weight between 25 and 170 kDa. Specificity and sensitivity of SCCWOS against serum pool from patients with chronic and acute forms of the disease were confirmed by immunoblot, which demonstrated that 25, 43, 60, 70, 85 and 160-kDa antigenic fractions of SCCWOS cultured for 5 and 10 days showed intense reactivity. We could demonstrate that SCCWOS of Pb are stable and show highly preserved antigenic fractions, which was proved by their high reactivity pattern to sera from different forms of PCM, anti-Pb antigen and anti-gp43 antisera.(AU)
Subject(s)
Paracoccidioides , Paracoccidioidomycosis , Serologic Tests , AntigensABSTRACT
The involvement of oxidization of peroxid in the antigen-antibody reaction makes the cell membrane permeability enhanced, so the fluorescence-labeled antibody stain can infiltrate into the cell easily and contact the antigen rapidly and diffusely, and then the antigen-antibody crosslinking can be formed efficiently. With the enhanced staining efficiency, shortened test duration, simplified operation, increased positive detection rate and accuracy, the new technology lays a foundation of clinical definite, curative effect view and prognosis.