ABSTRACT
The validation of kit of reagents destined to detection and quantitative evaluation of DNA of human cytomegalovirus in biological material using polymerase chain reaction technique in real time operation mode was implemented. The comparison was made against international WHO standard--The first WHO international standard for human cytomegalovirus to implement measures the kit of reagents "AmpliSens CMV-screen/monitor-FL" and standard sample of enterprise DNA HCMV (The central research institute of epidemiology of Rospotrebnadzor) was applied. The fivefold dilution of international WHO standard and standard sample of enterprise were carried out in concentrations of DNA HCMV from 106 to 102. The arrangement of polymerase chain reaction and analysis of results were implemented using programed amplifier with system of detection of fluorescent signal in real-time mode "Rotor-Gene Q" ("Qiagen", Germany). In the total of three series of experiments, all stages of polymerase chain reaction study included, the coefficient of translation of quantitative evaluation of DNA HCMV from copy/ml to ME/ml equal to 0.6 was introduced for this kit of reagents.
Subject(s)
Cytomegalovirus/genetics , DNA, Viral/genetics , Reagent Kits, Diagnostic , Real-Time Polymerase Chain Reaction , Humans , Real-Time Polymerase Chain Reaction/instrumentation , Real-Time Polymerase Chain Reaction/methods , Real-Time Polymerase Chain Reaction/standards , Sensitivity and SpecificityABSTRACT
Cerebral toxoplasmosis is one of the leading causes of neurologic diseases with high mortality rates in patients with HIV infection. Invasion was difficult to diagnose for a number of objective reasons. The objective of the investigation was to determine the clinical sensitivity of different laboratory techniques as both a single study and their various combinations to verify the diagnosis of cerebral toxoplasmosis in HIV-infected patients. Blood and cerebrospinal fluid were tested in 51 patients with Stage 4B HIV infection (AIDS) with the verified diagnosis of cerebral toxoplasmosis. Separate determination of specific antibodies of IgG, IgM, IgA and toxoplasma DNA in the blood and cerebrospinal fluid was shown to have an insufficient clinical sensitivity (37.3-68.6%). The benefits of various combinations of immunological and molecular biological assays enhancing the diagnostic efficiency up to 76.5-96.1% are demonstrated.
Subject(s)
Antibodies, Protozoan/blood , Brain/pathology , DNA, Protozoan/blood , HIV Infections/pathology , HIV , Toxoplasma/isolation & purification , Toxoplasmosis, Cerebral/diagnosis , Adult , Antibodies, Protozoan/cerebrospinal fluid , Brain/parasitology , Brain/virology , Coinfection , DNA, Protozoan/cerebrospinal fluid , Disease Progression , Female , HIV Infections/blood , HIV Infections/cerebrospinal fluid , HIV Infections/virology , Humans , Immunoassay , Immunoglobulin A/blood , Immunoglobulin A/cerebrospinal fluid , Immunoglobulin G/blood , Immunoglobulin G/cerebrospinal fluid , Immunoglobulin M/blood , Immunoglobulin M/cerebrospinal fluid , Magnetic Resonance Imaging , Male , Middle Aged , Polymerase Chain Reaction , Toxoplasma/immunology , Toxoplasmosis, Cerebral/blood , Toxoplasmosis, Cerebral/cerebrospinal fluid , Toxoplasmosis, Cerebral/parasitologySubject(s)
DNA, Protozoan/blood , Host-Parasite Interactions , Toxoplasma/physiology , Toxoplasmosis, Animal/immunology , Administration, Oral , Animals , Antigens, CD/genetics , Antigens, CD/immunology , B-Lymphocytes/immunology , B-Lymphocytes/pathology , Cell Movement , Cytokines/blood , Cytokines/immunology , Dendritic Cells/immunology , Dendritic Cells/pathology , Flow Cytometry , Genetic Predisposition to Disease , Immunophenotyping , Injections, Intraperitoneal , Macrophages/immunology , Macrophages/pathology , Mice , Mice, Inbred BALB C , Mice, Transgenic , Neutrophils/immunology , Neutrophils/pathology , Survival Rate , T-Lymphocytes/immunology , T-Lymphocytes/pathology , Toxoplasma/pathogenicity , Toxoplasmosis, Animal/genetics , Toxoplasmosis, Animal/mortality , Toxoplasmosis, Animal/pathologyABSTRACT
Ninety children, aged from 7 days to 12 months, with perinatal damage of the central nervous system (CNS) were virologically tested. The persistence of cytomegalovirus (CMV) DNA in the blood, urine and oropharyngeal swabs was identified in 18.9% of children. In 14.4% of children, CMV DNA was found only in oropharyngeal swabs and urine samples. In some cases, the severity of neurologic symptoms and absence of positive changes in the restoration treatment could be caused by the activity of CMV infection, blood CMV DNA and the increase in blood-brain barrier permeability. The authors recommend performance of a comprehensive immunological and molecular-biological survey in patients with suspected CMV infection. In case of positive reactions, a specific antiviral treatment should be started. Tree clinical cases are reported.
