ABSTRACT
4 primer sets, were used to allow the amplification of a nucleotide sequence with unique size for each of the dengue virus serotypes by polymerase chain reaction (PRC). The method consisted in the extraction of ribonucleic acid from supernatant of infected cell cultures, reverse transcription-polymerase chain reaction (RT-PCR). This was completed in approximately 7 hours in a simple tube. The size of the amplified sequence was evidenced by electrophoresis in Agarose gel stained with ethidium. The method showed a sensitivity of at least 2.5 plate forming units (pfu) per tube of reaction. It es useful for the detection and simultaneous identification of the 4 serotypes, starting from supernatants of infected strains cultures from different countries of the Caribbean, Central America, and South America.
Subject(s)
Dengue Virus/classification , Base Sequence , DNA Primers , Dengue Virus/genetics , Dengue Virus/isolation & purification , Humans , Immunoblotting/methods , Molecular Sequence Data , Nucleic Acid Hybridization/methods , Polymerase Chain Reaction/methods , RNA, Viral/isolation & purification , Sensitivity and Specificity , Serotyping/methodsABSTRACT
We present the results of the in vitro action of alpha and gamma interferons and of Intacglobin and Igegam against the 47/93/IPK (Coxsackie A9) strain isolated from the cerebrospinal fluid of a patient with epidemic neuropathy. The in vitro studies showed that the two interferons inhibited the replication of this agent; they also showed the presence of antibodies to it in the Intacglobin and Igegam. The results attained demonstrated that the use of these compounds could be effective for the treatment of this entity.
Subject(s)
Antiviral Agents/pharmacology , Enterovirus/drug effects , Interferon-alpha/pharmacology , Interferon-gamma/pharmacology , Virus Replication/drug effects , Animals , Chlorocebus aethiops , Coxsackievirus Infections/cerebrospinal fluid , Coxsackievirus Infections/epidemiology , Cuba/epidemiology , Enterovirus/physiology , Humans , Vero Cells/drug effectsABSTRACT
An indirect ultramicro ELISA system (UME) for the determination of antibodies to measles virus by the cuban technology of Ultramicro Analytical Systems (SUMA) was developed. A number of 448 serum samples from the National System of Seroepidemiological Surveillance of Measles were simultaneously processed by the Ultramicro ELISA system and by the haemagglutination inhibition technique. Results were compared in both assays. Afterwards; 120 serum pairs obtained from the same source were studies and processed in a similar way. Results evidenced the usefulness of such immunoenzymatic system for the serological diagnosis and the positive validation of replacing conventional techniques by this novel technology.
Subject(s)
Antibodies, Viral/analysis , Enzyme-Linked Immunosorbent Assay/methods , Measles virus/immunology , Animals , Chlorocebus aethiops , Humans , Measles virus/physiology , Sensitivity and Specificity , Vero Cells , Virus ReplicationSubject(s)
Animal Population Groups/microbiology , Animals, Wild/microbiology , Arboviruses/isolation & purification , Animals , Antibodies, Viral/analysis , Arboviruses/immunology , Birds/microbiology , Cuba , Disease Reservoirs , Ecology , Encephalitis Virus, Eastern Equine/isolation & purification , Encephalomyelitis, Equine/microbiology , Encephalomyelitis, Equine/transmission , Humans , Mammals/microbiology , Reptiles/microbiology , Species SpecificityABSTRACT
El virus del dengue 2 fue aislado en 22 de 40 sueros de pacientes clinicamente diagnosticados como dengue durante la epidemia de dengue hemorragico de Cuba 1981.Se utilizaron como sistemas de aislamientos el raton lactante y las celulas LLCMK2 demostrandose la mayor sensibilidad de estas al obtener 13 aislamientos mas que en raton lactante. Todas las cepas fueron identificadas como dengue tipo 2
