RESUMO
The aim of this study was to compare the data obtained using the new LIAISON® XL chemiluminescence system to search for HBsAg, anti-HCV, and anti-HIV1-2/p24 Ag with those obtained using the VITROS system currently adopted by the Microbiology Unit of the Hospital of Legnano. Routine samples of patients who were referred by practitioners for the determination of HBsAg (1,000 samples) and/or anti-HCV (1,002 samples) and/or anti-HIV1-2 (995 samples) were simultaneously analyzed using both systems. The concordant positive and discordant samples were re-examined for confirmation by means of an HBsAg neutralization assay, anti-HCV immunoblot, or anti-HIV1-2 Western blot; HBV-DNA, or HCV-RNA or HIV-RNA was also sought in the discordant samples. Samples of patients known to be positive were tested (100 HBsAg positive, 100 anti-HCV positive, and 100 HIV 1-2 positive) as well throughout treatment, with viremia levels becoming undetectable after treatment. The HBsAg, anti-HCV, and anti-HIV1-2 concordance between the two systems in routine series was respectively 99.8%, 98.5% and 99.7%, and 100% for all markers in samples known positive. The various molecular biology and confirmatory tests of the discordant samples were all negative (except for one anti-HCV positive sample). Measure of Cohen's kappa coefficient for HBsAg, anti-HCV, and anti-HIV gave K values of respectively 0.992, 0.946, and 0.980. In conclusion, the performance of the LIAISON® XL system in the routine laboratory determination for all three markers was comparable with that of the VITROS system. J. Med. Virol. 89:489-496, 2017. © 2016 Wiley Periodicals, Inc.
Assuntos
Anticorpos Anti-HIV/sangue , Infecções por HIV/diagnóstico , Antígenos de Superfície da Hepatite B/sangue , Hepatite B/diagnóstico , Anticorpos Anti-Hepatite C/sangue , Hepatite C/diagnóstico , Testes Sorológicos/métodos , Humanos , Medições LuminescentesRESUMO
Some international guidelines recommend evaluating the need to confirm positive anti-hepatitis C virus (HCV) antibody screening results by means of a more specific antibody or molecular biology test on the basis of a screening threshold value (such as the sample signal/cut-off ratio) that can predict the positivity of additional antibody testing in at least 95% of cases. The aim of this study was to determine the threshold value of the DiaSorin LIAISON XL chemiluminescence test. Two hundred and twenty-eight routine laboratory samples that were chemiluminescence positive for anti-HCV antibodies but had different signal/cut-off ratios were assayed using immunoblotting, which indicated that 155 (68.0%) were positive, 40 (17.5%) were negative, and 33 (14.5%) were indeterminate. When the samples were divided on the basis of their signal/cut-off ratios, 95.5% of the samples with a ratio of ≥3.5 were positive as against 74.1% of the positive or indeterminate samples with a ratio of <3.5. Statistical analysis using Youden's index and a receiver operating characteristic curve showed that the optimum cut-off value was 3.65. These findings indicate that, when using the LIAISON XL system for anti-HCV antibody screening, a signal/cut-off ratio of ≥3.65 makes further confirmatory tests unnecessary.
Assuntos
Doadores de Sangue , Anticorpos Anti-Hepatite C/sangue , Hepatite C/diagnóstico , Imunoensaio , Immunoblotting , Medições Luminescentes/métodos , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Criança , Feminino , Hepacivirus/imunologia , Hepatite C/epidemiologia , Hepatite C/imunologia , Humanos , Itália , Masculino , Pessoa de Meia-Idade , Curva ROC , Estudos Retrospectivos , Adulto JovemRESUMO
The presence of VCA IgG in the absence of VCA IgM and EBNA-1 IgG antibodies makes classifying EBV infection more difficult as this serological picture can be seen in the case of past infection with EBNA-1 IgG loss or non-appearance, or acute infections with the early disappearance or delayed onset of VCA IgM. The aim of this study was to assess the prevalence of this pattern in 2,422 outpatients with suspected EBV infection examined in 2005-2006, and to interpret its significance by means of immunoblotting. One hundred and seventy-seven (7.3%) of the patients were VCA IgG-positive, VCA IgM-negative and EBNA-1 IgG-negative, 15 of whom (8.5%) presented with heterophile antibodies. Analysis by age class showed that the prevalence of isolated VCA IgG ranged from 4.5% in the subjects aged 1-10 years to 9% in those aged >60 years. Immunoblotting allowed 18.9% of the cases to be classified as acute and 81.1% as past infections, the latter being observed in about 37% of the patients aged less than 10 years and in 100% of those aged >30 years. Therefore, in our case series, the presence of isolated VCA IgG was associated usually with past infection, particularly among adults. In children aged less than 10 years, it was associated mainly with acute infection but as past infection may be present in about one-third of such children, this possibility should not be overlooked.
Assuntos
Anticorpos Antivirais/sangue , Infecções por Vírus Epstein-Barr/epidemiologia , Infecções por Vírus Epstein-Barr/virologia , Herpesvirus Humano 4/imunologia , Imunoglobulina G/sangue , Mononucleose Infecciosa/virologia , Adolescente , Adulto , Distribuição por Idade , Idoso , Idoso de 80 Anos ou mais , Anticorpos Antivirais/imunologia , Antígenos Virais/imunologia , Proteínas do Capsídeo/imunologia , Criança , Pré-Escolar , Infecções por Vírus Epstein-Barr/imunologia , Antígenos Nucleares do Vírus Epstein-Barr/imunologia , Feminino , Herpesvirus Humano 4/classificação , Herpesvirus Humano 4/isolamento & purificação , Humanos , Imunoglobulina G/imunologia , Lactente , Mononucleose Infecciosa/imunologia , Masculino , Pessoa de Meia-Idade , Estudos Soroepidemiológicos , Adulto JovemRESUMO
The fetal consequences of CMV infection make it one of the most serious infections contracted during pregnancy, but the scientific community is divided over the proposed implementation of preventive screening for anti-CMV antibodies. The aim of this study was to assess the incidence and risk of infection during pregnancy in 2817 women who underwent anti-CMV IgG and IgM antibody screening during the period 2005-2007. The prevalence of anti-CMV IgG antibodies was 68.3% (95% CI: 66.6-70.0); the seroconversion rate in the 892 seronegative women was 0.32%; the results of IgG avidity testing revealed an cumulative incidence of 1.4% (95% CI: 0.97-1.83), density incidence of 0.8% (as cases/pregnant woman-trimester) (95% CI: 0.47-1.13), and a risk of infection of 0.5% (95% CI: 0.24-0.76). The screening identified 13 cases of primary infection (84.6% of which occurred in the first trimester of pregnancy). The possibility to identify these cases and consequently to plan appropriate interventions, supports the use of screening during pregnancy, especially in the first trimester when the risk of infection is greater.
Assuntos
Anticorpos Antivirais/sangue , Infecções por Citomegalovirus/epidemiologia , Citomegalovirus/imunologia , Complicações Infecciosas na Gravidez/epidemiologia , Risco , Adolescente , Adulto , Infecções por Citomegalovirus/diagnóstico , Infecções por Citomegalovirus/virologia , Feminino , Humanos , Imunoglobulina G/sangue , Imunoglobulina M/sangue , Incidência , Itália , Pessoa de Meia-Idade , Gravidez , Complicações Infecciosas na Gravidez/diagnóstico , Complicações Infecciosas na Gravidez/virologia , Trimestres da Gravidez , Prevalência , Adulto JovemRESUMO
The severity of congenital Toxoplasma gondii infection underlines the need for a precise diagnosis of acute infection during pregnancy. The search for specific IgM has been widely used for this purpose, but their possible early disappearance or persistence over time limits their meaning. In order to estimate the positive predictive value of anti-Toxoplasma IgM testing, we made an epidemiological analysis of the presence of anti-Toxoplasma IgG and IgM using ELISA in 4786 subjects attending the Hospital of Legnano in 2004-2005: 1360 seen for a clinical check-up and 3426 pregnant women for serological screening. In relation to IgG avidity, the positive predictive value of IgM was 45.98% (95% CI: 35.51-56.45) as a whole: this increased to 83.87% (95% CI: 70.92-96.82) in the patients with a highly positive test for IgM, but decreased to 9.52% (95% CI: 0.00-22.07) in pregnant women with a weakly positive test for IgM. Our results indicate that a highly positive IgM value in patients can be a good index of recent infection, but its poor predictive value in pregnant women underlines the need for additional tests with a follow-up if necessary.
Assuntos
Anticorpos Antiprotozoários/sangue , Imunoglobulina M/sangue , Complicações Parasitárias na Gravidez/diagnóstico , Toxoplasma/imunologia , Toxoplasmose/diagnóstico , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Animais , Criança , Pré-Escolar , Ensaio de Imunoadsorção Enzimática , Feminino , Humanos , Imunoglobulina G/sangue , Lactente , Masculino , Pessoa de Meia-Idade , Valor Preditivo dos Testes , Gravidez , Complicações Parasitárias na Gravidez/epidemiologia , Complicações Parasitárias na Gravidez/imunologia , Complicações Parasitárias na Gravidez/parasitologia , Toxoplasmose/epidemiologia , Toxoplasmose/imunologia , Toxoplasmose/parasitologiaRESUMO
BACKGROUND: Simultaneous detection of HIV 1 and 2 antibodies and HIV-1 p-24 antigen in the 4th generation tests is particularly effective for the identification of early acute HIV infections while maintaining accurate detection of long-established infections. OBJECTIVES: The aim of this study was to evaluate the new 4th generation VITROS HIV Combo test from Ortho-Clinical Diagnostics by comparing its results with those obtained using a 3rd generation HIV 1/2 antibody test (VITROS Anti HIV 1 + 2 from Ortho-Clinical Diagnostics) and a 4th generation test (LIAISON XL HIV Ab/Ag, DiaSorin) currently used in the Microbiology Unit of Legnano Hospital. STUDY DESIGN: One thousand and three samples of the normal daily routine (Group 1) were analyzed simultaneously with the three systems. The concordant and discordant sample results were further tested using Western blot and HIV-RNA assay (Roche). One hundred samples (Group 2) of known HIV positive subjects (63 of subtype B, 37 subtype non-B, and 51 with positive viraemia) and 50 samples (Group 3) with indeterminate Western blot were also examined using the three systems. From Group 3, 24 samples were collected from patients diagnosed with acute infection. RESULTS: The overall agreement between the three systems was 99.4% (99.5% in group 1, 100% in group 2 and 96.6% in group 3) with a coefficient Fleiss Kappa of 0.9814. Notably, the VITROS HIV Combo test was positive in all known HIV positive samples of group 2 without any statistically significant difference in the values of the sample/cut off ratios between the B and non-B subtypes and between the positive and negative viraemia samples in established infections. The VITROS HIV Combo test was also positive in all samples of patients with acute infection in group 3. CONCLUSIONS: The VITROS HIV Combo test has shown comparable performance to the other two assays in use of 3rd and 4th generation tests and is able to correctly identify both acute and established HIV infections independently of viraemia and HIV subtype.
Assuntos
Anticorpos Anti-HIV/sangue , Infecções por HIV/diagnóstico , Técnicas de Diagnóstico Molecular/normas , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Criança , Pré-Escolar , Feminino , Anticorpos Anti-HIV/isolamento & purificação , Proteína do Núcleo p24 do HIV/isolamento & purificação , HIV-1/isolamento & purificação , HIV-2/isolamento & purificação , Humanos , Masculino , Programas de Rastreamento , Pessoa de Meia-Idade , Técnicas de Diagnóstico Molecular/métodos , Reação em Cadeia da Polimerase , RNA Viral/genética , Kit de Reagentes para Diagnóstico/normas , Sensibilidade e Especificidade , Testes Sorológicos , Viremia/diagnóstico , Adulto JovemRESUMO
BACKGROUND: The emergence of the plasmid-mediated colistin resistance mechanism in Escherichia coli has raised concern among public health experts as colistin is a last-line antimicrobial resort. The primary aim of the study was to investigate the prevalence of this resistance trait in E. coli isolates circulating in the Lombardy region, Northern Italy. The presence of mcr-type genes and their genetic relationship were also studied. MATERIALS AND METHODS: A prospective study was performed during a 4-month period (May to August, 2016) in six acute care Hospitals. Consecutive nonduplicate clinical isolates of E. coli from any type of clinical specimen, with the exception of rectal swabs, were included in the study. Isolates that exhibited MIC values for colistin >2 mg/L were further investigated. Bacterial identification was obtained by matrix-assisted laser desorption ionization-time of flight mass spectrometry. Amplification of mcr-type genes (-1 to -5 variants) and microarray analysis were accomplished. Repetitive sequence-based PCR (Rep-PCR) and multilocus sequence typing (MLST) analysis were used for genotyping. RESULTS: Overall, 3,902 consecutive E. coli isolates (2,342 from outpatients, 1,560 from inpatients) were evaluated during the study period. Of them, 18/3,902 (0.5%), collected from 4/6 centers, showed resistance to colistin. These isolates were mostly obtained from urine of both outpatients (n=12) and inpatients (n=6). Colistin MIC values ranged from 4 to 8 mg/L. The mcr-1 gene was detected in 10/18 isolates (7 from outpatients, 3 from inpatients). Rep-PCR and MLST analysis revealed the presence of nine different clusters. Further mcr-type genes were not detected. CONCLUSION: Resistance to colistin in E. coli clinical isolates appears low in our geographic area. With regard to mcr-1-positive isolates, they accounted for approximately 50% of colistin-resistant E. coli isolates, thus representing a relevant resistance mechanism in this context. Although overall limited, the presence of mcr-1 determinant in our region should not be ignored and great concern should be given to the continuous surveillance.
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In order to avoid the influence of pre-analytical steps, the following study was performed by using sterile blood spiked with defined loads of microorganisms as inoculum. Time-to-Detection (TTD) was evaluated for the most frequently encountered bacteria and fungi in septicemia, comparing two commercially available blood culture systems, BACTEC 9240 (Becton Dickinson) and BacT/ALERT (Organon Teknika). A specific medium, Bactec Mycosis IC/F (Becton Dickinson), was compared with the Bactec Plus Aerobic (Becton Dickinson) and FAN Aerobic (Organon Teknika) media for recovery of fungi in general and in case of mixed bacterial/fungal septicemia. The results show that the BACTEC system detects nearly all enrolled microorganisms significantly faster than the BacT/ALERT; the anaerobic vial contributes to the detection of anaerobes and facultative anaerobes and, in the case of BACTEC, shortens TTD; the Bactec Mycosis IC/F bottle shortens TTD of fungi.
Assuntos
Bactérias/crescimento & desenvolvimento , Técnicas Bacteriológicas , Sangue/microbiologia , Fungos/crescimento & desenvolvimento , Aerobiose , Anaerobiose , Bacteriemia/microbiologia , Bactérias/isolamento & purificação , Contagem de Colônia Microbiana , Meios de Cultura , Fungemia/microbiologia , Fungos/isolamento & purificação , Humanos , Kit de Reagentes para Diagnóstico , Fatores de TempoRESUMO
BACKGROUND: The implementation of mass vaccinations against hepatitis B virus (HBV) has significantly reduced the prevalence of HBsAg-positive subjects. At the same time, the prevalence of the other markers of infection has decreased, but there has been an increase in the percentage of subjects with markers of a successful vaccination. It has been suggested that increasing immigration from countries in which this virus is highly endemic is changing the epidemiology of HBV infection. The aim of this study was to assess the prevalence of the serological markers of HBV in Italian and non-Italian HBsAg-negative subjects. MATERIALS AND METHODS: In the years 2007-2008, 8,018 samples from HBsAg-negative subjects (7,521 Italians and 497 non-Italians) were received for detection of anti-HBs and/or anti-HBc. The findings in the 1,358 samples from candidate blood donors were compared with those obtained in 1991 and 1999. RESULTS: The rate of anti-HBc positivity was 18.3% in the Italian samples and 32.8% in the non-Italian samples; the corresponding percentages of anti-HBs/anti-HBc positive samples (indicating past infection), anti-HBs positive only samples (vaccination) and anti-HBc positive only were, 11.3% vs. 22.5%, 25.8% vs. 17.2%, and 6.9% vs. 9.9% in Italians and non-Italians, respectively. The differences were more marked when stratified by age. In relation to candidate blood donors, simultaneous positivity for anti-HBs and anti-HBc decreased from 11.0% in 1991 to 8.1% in 1999 and 3.9% in 2007-2008, whereas isolated anti-HBs positivity increased from 2.2% in 1991 to 21.4% in 1999 and 42.9% in 2007-2008. CONCLUSIONS: The frequency of markers of past infection among Italians has decreased over time as a result of mass vaccination and is significantly lower than that observed in non-Italians. The increasing number of immigrants from countries in which HBV is highly endemic is changing the epidemiology of HBV infection in Italy.
Assuntos
Antígenos de Superfície da Hepatite B/sangue , Vírus da Hepatite B , Hepatite B , Vacinação , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Criança , Pré-Escolar , Feminino , Hepatite B/sangue , Hepatite B/epidemiologia , Hepatite B/prevenção & controle , Vacinas contra Hepatite B , Humanos , Lactente , Itália/epidemiologia , Masculino , Pessoa de Meia-Idade , Prevalência , Estudos RetrospectivosRESUMO
Various countries have implemented anti-rubella vaccination campaigns with the main aim of preventing congenital infection. In 2003, Italy joined the European WHO programme for the elimination of congenital rubella and issued a special healthcare plan, one of the objectives of which was to reduce the proportion of rubella-susceptible pregnant women to less than 5% by 2005. The aim of this study was to determine the percentage of seronegative pregnant women after the implementation of this plan. Anti-rubella IgG and IgM antibodies were sought in 2385 pregnant women who attended our hospital for serological screening between 1 July 2008 and 30 June 2010. They included 750 women of foreign origin (31.4%). Eight percent of the women were anti-rubella seronegative: 6.2% of the Italians and 11.7% of the non-Italians. Among the women of foreign origin, the percentage of seronegativity ranged from 5.6% of those coming from Eastern Europe to 17.7% of those coming from Latin America. The level of seropositivity among women of Italian origin is high, although the objective of ensuring less than 5% of susceptible pregnant women has not yet been quite reached in our area. However, particular attention needs to be given to women coming from geographical areas characterised by different epidemiologies and vaccination strategies because the percentage of seronegativity is in some cases double that of Italian women.
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BACKGROUND: Since 1998, Italian law requires serological screening for toxoplasmosis by the thirteenth week of pregnancy, and seronegative women should undergo further checks every 30 - 40 days until delivery (a total of 5 - 7 screenings). This is an improvement of the previous law which foresaw three free tests (one by the end of the third month, one in the fifth, and one in the ninth month of pregnancy). The aim of this study was to assess the implementation, in an urban area of Northern Italy, of the 1998 law about 10 years after its entry into force. METHODS: Of the 4,694 women who initiated and completed a pregnancy in the period 2006 - 2008, we recorded the trimester of pregnancy in which they underwent their first screening, the total and average number of screenings during pregnancy, and the trimester distribution of the screenings. RESULTS: A total of 84.1% of the women underwent their first screening during the first trimester. The negative cases underwent an average of 3.7 screenings during pregnancy, with 34.9% undergoing five or more. Sixty percent of the women underwent at least one screening per trimester. CONCLUSIONS: Our data indicates active screening during the first trimester, but fewer screenings than required by law during pregnancy as a whole. Therefore further efforts are needed to improve screening implementation. KEYWORDS: Anti-Toxoplasma antibodies; Congenital infection; Pregnancy trimester; Toxoplasma screening; Seroconversion; Seronegative women; Seropositive women; Screening protocol.
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A search for specific IgM antibodies was used for the detection of primary human cytomegalovirus (HCMV) infections, but the significance of the results is limited by the possible persistence of specific IgM over time, the fact that they are also produced during episodes of reactivation or reinfection, and possible cross-reactions with other viruses. Anti-HCMV antibody screening was carried out to assess the positive predictive value of detecting specific IgM antibodies using IgG and IgM enzyme-linked immunosorbent assays (ELISAs) in 6990 patients examined during the period 2005-2007. In comparison with IgG avidity, the positive predictive value of screening by IgM ELISA alone was 49.3%, which increased to 73% when the presence of IgM was confirmed by an enzyme-linked fluorescent assay (ELFA). The predictive values of highly or weakly positive IgM ELISA alone were respectively 68.8% and 16.4%, but increased to 83.1% and 39.1% if IgM was confirmed by ELFA. The positive predictive value of the IgM/IgG ratio ranged from 26.7% for a low ratio and the detection of IgM by ELISA alone, to 90.7% for a high ratio and ELFA-confirmed IgM detection. These findings indicate that a specimen in which highly positive IgM ELISA values are confirmed by ELFA, or which shows a high IgM/IgG ratio, is a good indicator of infection occurring within the previous three months.
Assuntos
Anticorpos Antivirais/sangue , Infecções por Citomegalovirus/diagnóstico , Citomegalovirus/imunologia , Imunoglobulina M/sangue , Virologia/métodos , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Afinidade de Anticorpos , Criança , Pré-Escolar , Ensaio de Imunoadsorção Enzimática/métodos , Feminino , Humanos , Imunoglobulina G/sangue , Lactente , Masculino , Pessoa de Meia-Idade , Valor Preditivo dos Testes , Gravidez , Adulto JovemRESUMO
The presence of an "isolated viral capsid antigen (VCA) IgG" pattern in serum is not easy to interpret without the aid of further tests, such as specific immunoblotting or a virus genome search, that often give rise to organisational and economic problems. However, one alternative is to use an enzyme-linked immunosorbent assay (ELISA) to detect anti-early antigen (EA) antibodies, which can be found in about 85% of subjects with acute Epstein-Barr virus (EBV) infections. The purpose of this work was to search for anti-EA(D) antibodies in 130 samples with an isolated VCA IgG pattern at ELISA screening and classified as being indicative of past (102 cases) or acute (28 cases) infection on the basis of the immunoblotting results. Thirty-seven samples (28.5%) were positive for anti-EA(D), of which 25 (89.3%) had been classified by immunoblotting as indicating acute and 12 (11.8%) past EBV infection. This difference was statistically significant (P < .01). The results of our search for anti-EA(D) antibodies correctly identified nearly 90% of acute (presence) or past EBV infections (absence). When other tests are not available, the search for anti-EA antibodies may therefore be helpful in diagnosing patients with an isolated VCA IgG pattern at screening tests.