ABSTRACT
The aim of this study was to evaluate an enzyme-linked immunoassay with recombinant rhoptry protein 2 (ELISA-rROP2) for its ability to detectToxoplasma gondii ROP2-specific IgG in samples from pregnant women. The study included 236 samples that were divided into groups according to serological screening profiles for toxoplasmosis: unexposed (n = 65), probable acute infection (n = 48), possible acute infection (n = 58) and exposed to the parasite (n = 65). When an indirect immunofluorescence assay forT. gondii-specific IgG was considered as a reference test, the ELISA-rROP2 had a sensitivity of 61.8%, specificity of 62.8%, predictive positive value of 76.6% and predictive negative value of 45.4% (p = 0.0002). The ELISA-rROP2 reacted with 62.5% of the samples from pregnant women with probable acute infection and 40% of the samples from pregnant women with previous exposure (p = 0.0180). Seropositivity was observed in 50/57 (87.7%) pregnant women with possible infection. The results underscored that T. gondii rROP2 is recognised by specific IgG antibodies in both the acute and chronic phases of toxoplasmosis acquired during pregnancy. However, the sensitivity of the ELISA-rROP2 was higher in the pregnant women with probable and possible acute infections and IgM reactivity.
Subject(s)
Female , Humans , Pregnancy , Antigens, Protozoan/immunology , Enzyme-Linked Immunosorbent Assay/methods , Immunoglobulin G/blood , Membrane Proteins/immunology , Pregnancy Complications, Parasitic/diagnosis , Protozoan Proteins/immunology , Toxoplasmosis/diagnosis , Antigens, Protozoan/blood , Confidence Intervals , Enzyme-Linked Immunosorbent Assay/standards , Fluorescent Antibody Technique, Indirect , Immunoglobulin G/isolation & purification , Immunoglobulin M/blood , Immunoglobulin M/isolation & purification , Inventions/standards , Membrane Proteins/genetics , Predictive Value of Tests , Pregnancy Complications, Parasitic/blood , Pregnancy Complications, Parasitic/immunology , Protozoan Proteins/genetics , Recombinant Proteins , Reference Standards , Sensitivity and Specificity , Toxoplasma/immunology , Toxoplasmosis/blood , Toxoplasmosis/immunologyABSTRACT
A leptospirose é uma antropozoonose endêmica em todo o mundo, que afeta o homem e várias espécies de animais domésticos e silvestres. No início da infecção há produção de IgM para o controle da infecção e após alguns dias, IgG são produzidas e provocam lise das leptospiras circulantes. Objetivou-se neste estudo identificar depósitos de antígeno de leptospiras e imunoglobulinas no tecido renal, para avaliar o papel de imunoglobulinas na patogênese da nefropatia da leptospirose em suínos. Foram colhidas 139 amostras de sangue e rim de suínos das cidades de Teresina/PI e Timon/MA, que foram avaliadas pela SAM, imunoistoquímica e PCR. Nefrite intersticial, fibrose, vasculite, tumefação do tufo glomerular e hipercelularidade difusa foram as principais alterações histopatológicas encontradas. A imunoistoquímica detectou antígeno de leptospira em 60 suínos. Depósitos de IgG, IgM e IgA foram observados no endotélio de capilares glomerulares, dos capilares intertubulares e na cápsula de Bowman, com marcação focal, difusa, global e segmentar. A deposição de IgM e IgA foi significantemente maior nos suínos infectados. Estranhamente depósitos de IgG foi significantemente maior nos suínos não infectados, onde não havia presença de antígeno de leptospiras e nem lesão túbulo-intersticial. Concluímos que antígeno de leptospiras no rim de suínos está relacionado a depósitos de IgM e IgA mas não a depósitos de IgG.
Leptospirosis is an endemic worldwide anthropozoonosis, affecting humans and several species of domestic and wild animals. At the beginning of infection is the production of IgM to control the infection and after a few days, IgG is produced and cause lysis of circulating leptospires. The objective of this study was to identify deposits of immunoglobulins and antigens of leptospires in kidney tissue, to assess the role of immunoglobulins in the pathogenesis of leptospirosis nephropathy in pigs. We collected 139 blood samples and kidney of pigs from the cities of Timon/MA and Teresina/PI, to be evaluated by SAM, immunohistochemistry and PCR. Interstitial nephritis, fibrosis, vasculitis; swollen glomeruli hypercellularity and diffuse in a pig were main pathological changes found. Immunohistochemistry leptospira antigen detected in 60 pigs. Deposits of IgG, IgM and IgA were observed in the endothelium of glomerular capillaries, the capillaries intertubulares and the Bowman's capsule, marked focal, diffuse, global and segmental. The deposition of IgM and IgA was significantly higher in infected pigs, strangely deposits of IgG was significantly higher in non-infected pigs, where there was no presence of antigen leptospires nor tubulointerstitial injury. We conclude that Leptospira antigen in porcine kidney relates to deposits of IgM and IgA but not IgG deposits.
Subject(s)
Animals , Antigens/analysis , Swine Diseases/immunology , Immunohistochemistry/veterinary , Immunoglobulin A/isolation & purification , Immunoglobulin G/isolation & purification , Immunoglobulin M/isolation & purification , Leptospirosis/veterinary , Leptospira/isolation & purification , Kidney Diseases/veterinary , Polymerase Chain Reaction/veterinary , Serologic Tests/veterinaryABSTRACT
Purpose: Parvovirus B19 (B19) is associated with a wide range of diseases in humans, whose severity depends on the immunological and haematological status of the host. Objective: To determine the incidence of B19 DNA and specific IgM and IgG frequency among patients suffering from different haematological malignancies and to determine the viral load using real-time PCR. Materials and Methods: A total of 70 patients were included in the study, in addition to a control group consisting of 20 apparently healthy volunteers. B19 DNA quantitative analysis was performed using real-time PCR while screening for IgM and IgG anti-B19 antibodies was performed using ELISA. Results: B19 DNA was detected in 26 patients (36.14%) and 3 controls (15%) using real-time PCR. Anti-parvovirus B19 IgM antibodies were detected in 9 patients (12.6%) and 2 controls (10%). Anti-parvovirus B19 IgG antibodies were detected in 32 patients (45.71%) and 5 controls (25%). The difference between the patient and control groups was found to be statistically non-significant in all of the three tests (P < 0.05). The difference in B19 incidence among patients receiving multiple transfusions and non-transfused patients was also found to be statistically non-significant (P < 0.05). Conclusion: We found a high incidence of B19 infection among patients diagnosed with different types of haematological malignancies. We recommend that all cases of haematological disorders should be examined for specific antibodies and tested for the presence of B19 DNA in serum by PCR technique.
Subject(s)
Adult , Chi-Square Distribution , Human Experimentation , Humans , Immunoglobulin G/isolation & purification , Immunoglobulin M/isolation & purification , Neoplasms/diagnosis , Parvovirus B19, Human/isolation & purification , Real-Time Polymerase Chain Reaction/methods , Serologic Tests/methodsABSTRACT
Chikungunya (CHIK) fever is a re-emerging Aedes mosquito-transmitted viral disease caused by CHIK virus belonging to the Togaviridae family of genus Alphavirus. The disease is almost self-limiting, occurs with characteristic triad of sudden onset fever, rash and arthritis. During the recent outbreak CHIKV was also found to cause long-term arthralgia, severe neurological disease and even fatalities. Although there are no antiviral or vaccines available for CHIKV, still there are several advantages to diagnose the infection. The present article provides an overview of various diagnostic modalities available and its significance by searching PubMed MeSH terms "Chikungunya virus" and "Diagnosis" for recent articles. The gold standard of CHIKV diagnosis is culture, yet requires facilities and skills. Highly sensitive and specific PCR assays for CHIKV have been developed, but the reagents and equipment are costly for widespread use. Serological diagnosis by detecting IgM antibody is most widely used as it is relatively cheaper and easier to perform. Disadvantages of antibody testing are cross-reactivity with other alpha viruses, cannot differentiate between recent past and acute infection, and its sensitivity varies in clinical settings. When tested for diagnosing acute CHIKV disease, sensitivities were just 4 to 22% and after 1 week rose to more than 80%. As most acutely infected patients seek medical attention within the first few days of illness, the ideal test should detect RNA or antigen. Therefore, the more realistic aim would be to develop a reliable antigen detection assay that could be used in rural areas, where CHIKV infection often occurs.
Subject(s)
Alphavirus Infections/diagnosis , Chikungunya virus/analysis , Chikungunya virus/isolation & purification , Clinical Laboratory Techniques/instrumentation , Clinical Laboratory Techniques/methods , Diagnosis , Diagnostic Techniques and Procedures , Immunoglobulin M/isolation & purification , Polymerase Chain Reaction/methodsABSTRACT
BACKGROUND AND OBJECTIVES: Hantaviruses are rodent-borne viruses of the family Bunyaviridae that have been identified as aetiological agents of two human diseases, haemorrhagic fever with renal syndrome (HFRS) and hantavirus pulmonary syndrome (HPS). There are no reports of hantavirus infections in humans from India, hence this pilot study was undertaken to provide the serological evidence of hantavirus infections in humans in south India. METHODS: Serum samples were obtained from individuals with acute febrile illness and from voluntary blood donors, majority of whom were from south India. Serum samples were tested for anti-hantavirus IgM using a commercial enzyme immunoassay (EIA). Samples found positive by the EIA were tested by an indirect immunofluorescence assay (IFA) using slides coated with Seoul virus (SEOV) infected cells as substrate. RESULTS: Of the 152 serum samples from individuals with pyrexic illness, 23 (14.7%) were positive for anti-hantavirus IgM by EIA. In contrast, only 5.7 per cent of healthy blood donors were positive by this assay. Eighteen of the 22 (82%) EIA-positive samples from patients were positive by the IFA assay. In contrast, only 2 of the 5 (40%) blood donor EIA positive samples were positive in the IFA assay. INTERPRETATION AND CONCLUSION: The finding of this study indicated the possible presence of hantavirus infections in the human population of India presenting both as asymptomatic and symptomatic infections. Further studies need to be done to confirm the findings on a larger sample using molecular techniques.
Subject(s)
Fluorescent Antibody Technique, Indirect , Hantavirus Infections/epidemiology , Humans , Immunoenzyme Techniques , Immunoglobulin M/isolation & purification , India/epidemiology , Pilot Projects , Serologic Tests/methodsABSTRACT
Considerando que algunos autores han reportado un aumento en la cantidad de algunas inmunoglobulinas en los pacientes con actinomicetoma, en este trabajo nos propusimos determinar diferencias en la producción de IgG1, IgG2, IgG3, IgG4 e IgM en 25 pacientes con actinomicetoma por Nocardia brasiliensis y 25 personas sanas provenientes de una zona endémica de micetoma. La determinación de inmunoglobulinas se realizó por medio de la técnica de ELISA. Para sensibilizar las placas se emplearon 6 antígenos de N. brasiliensis: un antígeno crudo denominado NB y cinco derivados del mismo (NB2, NB4, NB6, NB8 y NB10) separados por punto isoeléctrico. Los niveles de las cuatro subclases de IgG fueron mayores en los sueros de los pacientes que en el suero de los controles, con una diferencia máxima en IgG3 e IgG4; para esta última subclase, los seis antígenos fueron altamente reactivos. La concentración de IgM fue igual en ambos grupos. Es probable que como ocurre en otras infecciones, en la fisiopatogenia del actinomicetoma influya no sólo el aumento o deficiencia de una clase de inmunoglobulina, sino la relación que existe entre las diferentes subclases.
Considering that some authors have reported an increasing of some immunoglobulins in actinomycetoma patients, in this study we propose to determine differential production of IgG1, IgG2, IgG3, IgG4 and IgGM in 25 patients with actinomycetoma and 25 healthy individuals from a mycetoma endemic area. Immunoglobulins were determined by ELISA technique. To sensibilize the plates, six Nocardia brasiliensis antigens were used: a crude antigen denominated NB and five derivatives (NB2, NB4, NB6, NB8 and NB10) obtained by their isoelectric point. Results showed that all IgG subclasses were higher in the patients’ sera than in control sera, with a maximal difference to IgG3 and IgG4. To the latter subclass, six antigens were highly reactives. IgM levels were similar in both groups. As it occurs in other infections, in the actinomycetoma pathogenesis probably participate the increase or deficiency of a determined immunoglobulin class, as well as the relationship between different subclasses.
Subject(s)
Adult , Female , Humans , Male , Antibodies, Bacterial/immunology , Mycetoma/immunology , Nocardia Infections/immunology , Antibody Specificity , Antibodies, Bacterial/blood , Antibodies, Bacterial/isolation & purification , Antigens, Bacterial/immunology , Enzyme-Linked Immunosorbent Assay , Isoelectric Point , Immunoglobulin G/blood , Immunoglobulin G/classification , Immunoglobulin G/immunology , Immunoglobulin G/isolation & purification , Immunoglobulin M/blood , Immunoglobulin M/immunology , Immunoglobulin M/isolation & purification , Mycetoma/microbiology , Nocardia Infections/bloodABSTRACT
Parvovirus B19 has been associated by some investigators with cases of severe hepatitis. The aim of the present study was to determine the presence of active parvovirus B19 infection among 129 Brazilian patients with non-A-E hepatitis. The patients were assayed for antibodies against parvovirus B19, IgM class, by ELISA. In IgM-positive cases, parvovirus B19 DNA was assayed by PCR in serum and liver tissue and parvovirus VP1 antigen in liver tissue was assayed by immunohistochemistry. Antibodies against parvovirus B19, IgM class, were detected in 3 (2.3 percent) of 129 patients with non-A-E hepatitis. Previous surgery and blood transfusions were reported by these 3 patients. One patient was a 56-year-old female with severe hepatitis, with antimitochondrial antibody seropositivity and submassive necrosis at liver biopsy, who responded to corticosteroid therapy. Strong evidence for active parvovirus B19 infection was found in this patient, with parvovirus B19 DNA being detected by PCR in liver tissue. Furthermore, parvovirus VP1 antigen was also detected in liver tissue by immunohistochemistry. The other two IgM-positive patients were chronic hepatitis cases, but active infection was not proven, since neither viral DNA nor antigen were detected in their liver tissues. This and other reports suggest a possible relation between parvovirus B19 infection and some cases of hepatitis
Subject(s)
Humans , Male , Female , Middle Aged , Hepatitis, Viral, Human/virology , Parvovirus B19, Human/isolation & purification , Acute Disease , Antibodies, Viral/isolation & purification , Antigens, Viral/isolation & purification , Chronic Disease , DNA, Viral/isolation & purification , Electrophoresis, Agar Gel , Enzyme-Linked Immunosorbent Assay , Immunoglobulin M/isolation & purification , Liver/pathology , Liver/virology , Parvovirus B19, Human/immunology , Polymerase Chain ReactionABSTRACT
A crude antigenic preparation of Babesia bigemina was used to develop an ELISA for the detection of IgM antibodies. Optimal dilutions of the antigen, using positive and negative reference sera, were determined by checkerboard titrations. Negative sera from cattle imported from tick-free areas, serum samples collected from infected B. bigemina cattle were used to validate the test. The specificity was 94 percent and sensitivity of the Elisa 87.5 percent. Sera from 385 cattle deriving from areas free from tick-borne diseases, which were submitted to a preimmunization process, were screened by this technique. The Elisa detected seroconversion on the 14th day post-inoculation in animals either infested with Boophilus microplus ticks (infected with B. bigemina), or inoculated with B. bigemina infected blood. Antibody titers decreased after day 33; however, all animals remained positive until the end of the experiment (124 days). The ELISA described may prove to be an appropriate serological test for the detection of IgM antibodies against B. bigemina
Subject(s)
Animals , Cattle , Babesia/immunology , Immunoglobulin M/isolation & purification , Babesiosis/blood , Babesiosis/diagnosis , Enzyme-Linked Immunosorbent Assay/methods , Immunoglobulin M/blood , Sensitivity and SpecificityABSTRACT
A prospective study of cytomegalovirus (CMV) infection was carried out on 34 renal transplant recipients managed at a General Hospital in Ribeirão Preto, SP, Brazil. Serologic tests showed that all patients were infected with CMV before renal transplantation. Two nested-PCR techniques with primers that recognize sequences of the glycoprotein B (gB) and H (gH) genes were used for CMV detection in blood and urine samples during the post-transplantation period. CMV was detected more frequently in blood samples than in urine samples (P<0.001). Thirty-three patients had CMV detected at least once in blood and/or urine samples. Seven of these patients (21.2 percent) were diagnosed as having symptomatic CMV infection and showed a worse clinical outcome, with a higher death rate (P = 0.03). No association between CMV viremia and graft rejection was observed. Nested-PCR was not useful to identify patients at risk for symptomatic CMV infection since only 21.2 percent of the patients with CMV infection were symptomatic
Subject(s)
Humans , Cytomegalovirus Infections/diagnosis , Kidney Transplantation , Polymerase Chain Reaction/methods , Base Sequence , Cytomegalovirus Infections/blood , Cytomegalovirus Infections/urine , DNA Primers , Immunoglobulin G/isolation & purification , Immunoglobulin M/isolation & purification , Prospective Studies , Viral Envelope Proteins/geneticsABSTRACT
Viral antigens for 4 dengue serotypes were produced in C6/36 Aedes albopictus cells. These were used as assay antigens for IgM-capture ELISA to detect IgM antibodies in sera of dengue patients from 3 hospitals in Metro Manila, Philippines. A total of 378 serum samples came from National Children's Hospital (NCH), San Lazaro Hospital (SLH), and St Luke's Medical Center (SLMC), from January to November 1995. Three hundred and four (304) out of 378 serum samples, or 80.42% showed positive IgM ELISA titer against at least one of the 4 assay antigens. Dengue type 4 (D4) antigen detected antibodies in 61.90% (234/378) of these serum samples, whereas type 1 (D1), type 3 (D3), and type 2 (D2) had detection rates of 60.05% (227/378), 50.79% (192/378) and 49.47% (187/378) respectively. Although the results show that both D1 and D4 are the most effective antigens in identifying dengue infections for this batch of samples, the use of a cocktail of antigens is still recommended. The results of this study are the basis for the IgM-capture ELISA protocol presently applied for the laboratory confirmation of dengue cases in the Philippines.
Subject(s)
Animals , Antibodies, Viral/blood , Dengue/blood , Enzyme-Linked Immunosorbent Assay/methods , Humans , Immunoglobulin M/isolation & purification , Insect Vectors , PhilippinesABSTRACT
The possibility of detecting acute infection and immunity using body fluids that are easier to collect than blood, mainly in children, would facilitate the investigation and follow-up of outbreaks of hepatitis A (HAV). Our study was carried out to evaluate the detection of anti-HAV IgM, IgA and total antibodies in saliva using serum samples as reference. Forty three paired serum and saliva samples were analyzed. From this total, 24 samples were obtained from children and 1 from one adult during the course of acute hepatitis A; an additional 18 samples were obtained from health professionals from Adolfo Lutz Institute. The sensitivity to detect anti-HAV IgM was 100 per cent (95 per cent CI: 79.1 to 100.0 per cent), employing saliva as clinical samples. In detecting anti-HAV IgA, the sensitivity was 80.8 per cent (95 per cent CI: 60.0 to 92.7 per cent) and for the total antibodies was 82.1 per cent (95 per cent CI: 62.4 to 93.2 per cent). The specificity was 100 per cent for each. The rate of agreement was high comparing the results of serum and saliva samples for detecting HAV antibodies. We conclude that saliva is an acceptable alternative specimen for diagnosing acute hepatitis A infection, and for screening individuals to receive hepatitis A vaccine or immunoglobulin.
Subject(s)
Humans , Child , Adult , Hepatitis Antibodies/isolation & purification , Hepatitis A/immunology , Saliva/immunology , Enzyme-Linked Immunosorbent Assay , Hepatitis Antibodies/blood , Hepatitis A/diagnosis , Hepatitis A/prevention & control , Immunoglobulin A/isolation & purification , Immunoglobulin M/isolation & purification , Saliva/virology , Sensitivity and SpecificityABSTRACT
We describe the production of the potential monoclonal antibodies (MoAbs) using BALB/c mice immunized with vesicular fluid (VF)-Tcra (T. crassiceps) antigen. Immune sera presented anti-VF-Tcra (<20kD) IgG and IgM antibodies with cross-reactivity with T. solium (Tso) antigen (8-12, 14, and 18 kD). After cell fusion, we selected 33 anti-Tcra and anti-Tso reactive IgM-clones and 53 anti-Tcra specific IgG-clones, 5 of them also recognizing Tso antigens. Two clones identified the 8-14 and 18kD peptides of VF-Tcra.
Subject(s)
Animals , Female , Mice , Antibodies, Helminth/biosynthesis , Antibodies, Monoclonal/biosynthesis , Antigens, Helminth/immunology , Taenia/immunology , Cross Reactions , Cysticercosis/immunology , Immunoblotting , Immunoglobulin G/immunology , Immunoglobulin G/isolation & purification , Immunoglobulin M/immunology , Immunoglobulin M/isolation & purification , Mice, Inbred BALB CABSTRACT
An ELISA test for trichinosis using as antigen a larvae soluble fraction from trichinella spiralis was carried out for the detection of IgM and IgA specific antibodies in 45 serum samples from patients confirmed or suspected to have trichinosis by strong clinical and epidemiological evidences. All the patients had positive serology detected by precipitin test, bentonite floculation test, indirect hemagglutination tes and ELISA IgG test. The cut off value was determined using two criteria. Criterion A was determined in each plate, using three positive controls and two negative ones; the average of the negative controls and the weakest positive control, muliplied by a 1,2 factor was, considered the cut off value. Criterion B was determinated using the average plus three standard deviations from 64 apparently halthy persons serum samples. In both cases, three serum dilutions (1:10, 1:100 and 1:500) were used. The sensitivity of ELISA IgM was 100,0, 93,3 and 82,2 percent using serum dilution of 1:10, 1:100 and 1:500 respectively (criterion A) and 100,0, 97,8 and 95,6 percent for the same dilutions (criterion B), whereas the values for ELISA IgA were: 100,0 91,1 and 86,7 percent (criterion A) and 100,0 100,0 and 91,1 percent (criterion B). In order to find out the specificity of ELISA IgM and ELISA IgA, additional 118 serum samples from individuals with other parasitoses, such as cysticercoss (18) hydatidosis (39), fascioliasis (12), toxocariasis (30), Chagaïs disease (12) and individuals with non specif eosinophilia (7), were also tested. ELISA IgM presentes a specificity of 92,3, 93,4 and 97,3 percent (criterion A) and 96,2, 97,8 and 97,8 percent (criterion B) whereas the results for ELISA IgA were 97,8, 98,9 and 99,4 percent (criterion A) and 98,4 percent for the 1:10 and 1:100 dilutions and 100,0 percent for the 1:500 dilution (criterion B). The positive predictive values of ELISA IgM were 76,3, 77,8 and 88,1 percent (criterion A) and 86,5, 91,7 and 91,5 percent (criterion B) whereas the negative ones were 100,0, 98,3 and 95,7 percent (criterion A) and 100,0, 99,4 and 98,9 percent (criterion B). The positive predictive values of ELISA IgA were 91,8, 95,3 and 97,5 percent (criterion A) and 93,8, 93,8 and 100,0 percent (criterion B) whereas the negatives ones were: 100,0, 97,9 and 96,8 percent (criterion A) and 100,0, 100,0 and 97,8 percent (criterion B). The use of ELISA IgM and ELISA IgA in the inmunodiagnosis of trichinosis is discussed
Subject(s)
Humans , Enzyme-Linked Immunosorbent Assay/standards , Trichinellosis/diagnosis , Immunoglobulin A/isolation & purification , Immunoglobulin M/isolation & purification , Sensitivity and SpecificityABSTRACT
To assess the clinical relevance of a semi-quantitative measurement of human cytomegalovirus (HCMV) DNA in renal transplant recipients within the typical clinical context of a developing country where virtually 100 per cent of both receptors and donors are seropositive for this virus, we have undertaken HCMV DNA quantification using a simple, semi-quantitative, limiting dilution polymerase chain reaction (PCR). We evaluated this assay prospectively in 52 renal transplant patients from whom a total of 495 serial blood samples were collected. The samples scored HCMV positive by qualitative PCR had the levels of HCMV DNA determined by end-point dilution-PCR. All patients were HCMV DNA positive during the monitoring period and a diagnosis of symptomatic infection was made for 4 of 52 patients. In symptomatic patients the geometric mean of the highest level of HCMV DNAemia was 152,000 copies per 106 leukocytes, while for the asymptomatic group this value was 12,050. Symptomatic patients showed high, protracted HCMV DNA levels, whereas asymptomatic patients demonstrated intermittent low or moderate levels. Using a cut-off value of 100,000 copies per 106 leukocytes, the limiting dilution assay had sensitivity of 100 per cent, specificity of 92 per cent, a positive predictive value of 43 per cent and a negative predictive value of 100 per cent for HCMV disease. In this patient group, there was universal HCMV infection but relatively infrequent symptomatic HCMV disease. The two patient groups were readily distinguished by monitoring with the limiting dilution assay, an extremely simple technology immediately applicable in any clinical laboratory with PCR capability.
Subject(s)
Humans , Cytomegalovirus , Cytomegalovirus Infections/diagnosis , Kidney Transplantation , Leukocytes/virology , Polymerase Chain Reaction , Viral Load , DNA , Immunoglobulin G/isolation & purification , Immunoglobulin M/isolation & purification , Prospective StudiesABSTRACT
Um total de 143 soros de pacientes (120 da forma cutânea localizada, seis da mucocutânea e 17 com leishmaniose visceral), provenientes de ambulatórios ou de hospitais do Grande Rio de Janeiro, suspeitos de leishmaniose tegumentar ou visceral americanas, foi submetido às reaçöes de imunofluorescência indireta (RIFI-IgM e IgG). Estes soros foram selecionados porque se apresentavam com RIFI-IgG de títulos elevados ou eram RIFI-IgM reagentes no soro. Como existe a possibilidade de falsos resultados de IgM näo reagentes na presença de títulos elevados de IgG e falsos IgM reagentes, devido à presença de fator reumatóide (autoanticorpos IgM anti-IgG), utilizou-se a separaçäo das fraçöes IgM e IgG do soro destes pacientes. Para isro, procedeu-se a separaçäo destas imunoglobulinas em coluna de Sephacryl S-300, nos casos em que os soros eram IgM negativo e IgG maior ou igual a 360, com a finalidade de se detectar falsos negativos e, em soros IgM reagentes, falsos positivos. Nestes últimos, também realizou-se a prova do látex para fator reumatóide. Deste modo a RIFI-IgM da fraçäo IgM separada no Sephacryl permitiu evidenciar apenas um soro - de paciente da forma cutânea localizada (0,7 por cento) - falso negativo por provável competiçäo com títulos de anticorpos IgG elevados. Por outro lado, permitiu o encontro de 23 (16,1 por cento) soros RIFI-IgM falso-reagentes para leishmanioses, devido à presença de fator reumatóide (seis soros eram de leishmaniose mucocutânea e os 17 restantes de leishmaniose visceral). Em outros indivíduos da forma cutânea localizada (8,4 por cento) a RIFI-IgM do soro e a RIFI da fraçäo IgM eram reagentes, embora também tivessem positivos (maior ou igual a 20 U/ml) à prova do látex, como os outros 23 indivíduos anteriores. Os 107 indivíduos restantes (74,8 por cento) foram RIFI-IgM reagentes no soro e na fraçäo IgM, mas, entretanto, todos eram negativos para fator reumatóide. Todas as RIFI-IgM das fraçöes IgM eram näo-reagentes. Os resultados demonstram a utilidade da presente metodologia na obtençäo de maior confiabilidade nos testes de RIFI-IgM para leishmanioses
Subject(s)
Humans , Chromatography, Gel , Fluorescent Antibody Technique, Indirect , Immunoglobulin M/isolation & purification , Latex Fixation Tests , Leishmaniasis, Cutaneous , Leishmaniasis, Diffuse Cutaneous , Leishmaniasis, Mucocutaneous , Leishmaniasis, Visceral , Rheumatoid Factor , False Negative Reactions , False Positive ReactionsABSTRACT
En este trabajo se presenta la normalización del procedimiento de detección de anticuerpos IgM contra el virus del dengue en muestras de sangre tomadas en papel de filtro. Las muestras se obtuvieron de 118 pacientes, de los cuales 91 tenían un diagnóstico clínico de dengue y 27 un diagnóstico de una infección viral que no guardaba relación con esa enfermedad, siendo los primeros originarios de Costa Rica y Nicaragua y de Cuba los segundos. En todas las muestras se determinó la presencia de anticuerpos IgM contra el virus del dengue mediante un inmunoensayo enzimático (ELISA) de captura. Al analizarse en su conjunto los resultados de los pacientes de los tres países se determinaron una sensibilidad y especificidad de 98,1 por ciento y 98,5 por ciento respectivamente, para la prueba efectuada con sangre entera en papel de filtro conservada a 4 grados centígrados, y una concordancia de 96 por ciento entre los resultados de esa prueba y los del ELISA. Cuando se compararon los resultados obtenidos con las tres muestras de un mismo paciente - las de sangre en papel de filtro conservadas a la temperatura ambiental y a 4 grados centígrados, y la del suero correspondiente - también se obtuvo una concordancia de 86 por ciento. Este estudio demuestra la elevada sensibilidad y especificidad diagnósticas logradas con el procesamiento de sangre entera absorbida en papel de filtro en las condiciones detalladas en el artículo. Los autores recomiendan utilizar este métodos en los programas de vigilancia de dengue en la Región
This paper presents a standardized procedure for the detection of IgM antibodies to dengue virus in blood samples taken from filter paper. The samples were obtained from 118 patients, of whom 91 had been clinically diagnosed with dengue and 27 with a viral infection unrelated to that disease. The first group of patients came from Costa Rica and Nicaragua and the second group from Cuba. All the samples were tested for IgM antibody against dengue virus by means of a capture enzyme immunoassay (EIA). Analysis of the results for patients from all three countries together yielded a sensitivity of 98.1% and a specificity of 98.5% for the test done on whole blood on filter paper stored at 4 °C; agreement between the results of that test and those of the EIA using serum samples was 96%. In a comparison of the results obtained with three samples from the same patientwhole blood on filter paper stored at room temperature, the same type of sample stored a 4 °C, and serumthe agreement was 86%. This study demonstrates the high diagnostic sensitivity and specificity achieved when whole blood absorbed on filter paper is processed in the manner described in detail in the article. The authors recommend the use of this method in the dengue surveillance programs in the Region.
Subject(s)
Paper , Immunoglobulin M/isolation & purification , Dengue Virus , Diagnosis , Blood Specimen Collection , Immunoenzyme Techniques , Epidemiological Monitoring , Costa Rica , Cuba , NicaraguaABSTRACT
Os autores apresentam o caso de uma paciente com 34 anos de idade, com cinco eventos de abordamento, fetal, livedo reticular, acidente vascular cerebral e positividade para os anticorpos anticardiolipina idiotipos IgG e IgM. A paciente preenche os critérios par síndrome Sneddon, associado às manifestaçoes clínicas e positividade para os anticorpos anticardiolipina. Chama-se atençao para a investigaçao dos anticorpos anticardiolipina na síndrome de Sneddon.
Subject(s)
Humans , Female , Adult , Antibodies, Anticardiolipin/isolation & purification , Sneddon Syndrome/immunology , Immunoglobulin G/isolation & purification , Immunoglobulin M/isolation & purification , Skin Diseases, Vascular/diagnosisABSTRACT
Se analizaron los datos con relación a la prevalencia de anticuerpos IgG e IgM contra el virus de la hepatitis A (VHA), mediante la técnica de ELISA en 450 ninos sin antecedentes de hepatitis, con edades comprendidas entre los 3 meses y 17 años de edad, que acudieron a consulta al Instituto Nacional de Pediatría de la ciudad de México en el período comprendido de septiembre de 1992 a junio de 1993. La prevalencia de anticuerpos IgG en la población estudiada, fue del 83.6 por ciento. De los niños menores de un año el 50 por ciento mostraron anticuerpo, el 80 por ciento a los 3 años 80 por ciento, y el 96 por ciento a los 10 años de edad. Sólo 9 niños de los 450 tuvieron además anticuerpos IgM contra el virus de la hepatitis A. Se concluye que la prevalencia de HVA en la población que estudiamos es semejante a lo referido hace 13 años en México.
Subject(s)
Humans , Male , Female , Infant , Child, Preschool , Child , Adolescent , Hepatitis Antibodies/isolation & purification , Hepatitis A/epidemiology , Hepatovirus/immunology , Enzyme-Linked Immunosorbent Assay , Epidemiology, Descriptive , Immunoglobulin G/isolation & purification , Immunoglobulin M/isolation & purification , Longitudinal Studies , Prevalence , Prospective StudiesABSTRACT
Relizou-se um estudo controlado para comparar a presença parasitemiaa e anticorpos antiplasmódios em doadores expostos ao risco de infecçäo malárica, segundo os critérios fixados nas Normas Técnicas em Hemoterapia, do Miníterio da Saúde. Eles estabelecem a rejeiçäo dos candidatos à doaçäo que apresentaram quadro febril há 30 dias e malária há 12 meses, ou que frequentaram área de malária há 6 meses. Foram estudados 395 candidatos incluídos nos critérios de rejeiçäo (expostos) e 383 candidatos aptos (controles), selecionados na triagem de doadores do Hemocentro de Manaus (AM). Em ambos os grupos, realizou-se a gota espessa e o teste da laranja acridina (QBC) para o diagnóstico parasitológico e a imunofluorescência indireta para a pesquisa de anticorpos IgM e IgG ao Plamodium vivax e falciparum. Observou-se diferença significante entre expostos e controles em relaçäo à presença de parasitemia, porém mäo para a presença de anticorpos antiplasmódios (P<0.05). O QBC foi mais sensível do que a gota espessa para detectar parasitemia e a concordância entre ambos foi de 0.66 ao Indice Kappa
Subject(s)
Humans , Male , Adult , Antibodies, Protozoan/isolation & purification , Blood Banks/standards , Blood Donors , Malaria/diagnosis , Endemic Diseases , Immunoglobulin G/isolation & purification , Immunoglobulin M/isolation & purification , Malaria/immunology , Plasmodium falciparum , Plasmodium vivax , Risk FactorsABSTRACT
We report five patients with vasculitis and antineutrophil cytoplasmic antibodies with cytoplasmic pattern. All had severe upper and lower respiratory tract necrotizing lesions. Three had kidney failure due to rapidly progressive glomerulonephritis. The pathological study showed a crescentic glomerulonephritis, a chronic granulomatous inflammation in the lungs and in the nasal mucosa, an acute nonspecific inflammation or a chronic granulomatous inflammation and focal blood vessel fibrinoid necrosis. All patients with simultaneous involvement of lungs and kidneys had high titers of antineutrophil cytoplasmic antibodies. The nomenclature and classification of these diseases is discussed