ABSTRACT
BACKGROUND: The 2013-2016 Ebola virus disease (EVD) outbreak showed a lack of diagnostic point-of-care methods. Currently, EBOV diagnosis relies on quantitative reverse-transcription-PCR (RT- qPCR), highly specific and sensitive, but requiring skilled personnel and well-equipped laboratories. In field settings, these factors and others, such as samples' time of collection and transportation, determine a prolonged turnaround-time to final results. In outbreak scenarios, a rapid and transportable method could eliminate issues of cohorting suspected and actual EVD patients for lack of diagnostic certainty. The aim of this study was the field evaluation of the new fast, easy-to-use and reliable RT-qPCR assay and platform for EBOV detection, developed in the framework of the EbolaMoDRAD project by CLONIT S.r.l. and STMicroelectronics S.r.l. STUDY DESIGN: We evaluated its performance during the outbreak and in further studies in the EVD laboratory at the Princess Christian Maternity Hospital (PCMH) in Freetown (Sierra Leone) run by Emergency NGO and the Italian National Institute for Infectious Diseases (INMI). The assay was tested on residual aliquots of clinical specimens from EBOV-positive or -negative patients (n=116, EVD prevalence 37%). RESULTS AND CONCLUSION: Overall, the test was very easy-to-use and the instrument was robust and reliable in field-settings. The sensitivity of the assay was 100% and the specificity was 98.63% (95%CI: 96.34-100.92%). The positive and negative predictive values were 97.73 (95%CI:94.77-100.68%) and 100%, respectively. The high sensitivity and specificity of this new assay indicate that it is promising for laboratory diagnosis, especially in resource-limited settings.
Subject(s)
Ebolavirus/isolation & purification , Hemorrhagic Fever, Ebola/diagnosis , Point-of-Care Systems , Real-Time Polymerase Chain Reaction/methods , Adult , Disease Outbreaks/prevention & control , Female , Humans , Male , Prevalence , RNA, Viral/analysis , Sensitivity and Specificity , Sierra LeoneABSTRACT
We present a clinical case of antiretroviral treatment failure with appearance of mutations demonstrated by genotyping. We also show the evolution of the pattern of mutations that confers resistance to protease and reverse transcriptase inhibitors along with changes in the scheme of drugs indicated to the patient. A deletion was found in codon 67 of the TR gen, along with a novel resistance model to AZT pointing out the benefits of the detection of antiviral resistance by sequencing (genotyping).
Subject(s)
Acquired Immunodeficiency Syndrome/drug therapy , Anti-HIV Agents/therapeutic use , Gene Deletion , RNA-Directed DNA Polymerase/genetics , Zidovudine/therapeutic use , Adult , Amino Acid Sequence , Base Sequence , Drug Resistance, Microbial/genetics , Female , Genotype , HIV Protease/genetics , Humans , Treatment FailureABSTRACT
We present a clinical case of antiretroviral treatment failure with appearance of mutations demonstrated by genotyping. We also show the evolution of the pattern of mutations that confers resistance to protease and reverse transcriptase inhibitors along with changes in the scheme of drugs indicated to the patient. A deletion was found in codon 67 of the TR gen, along with a novel resistance model to AZT pointing out the benefits of the detection of antiviral resistance by sequencing (genotyping).
ABSTRACT
En la práctica diaria, cuando se realiza la determinación de creatinquinasa isoenzima MB (CK-MB) por el método inmunológico, es habitual obtener resultados incongruentes: valores de CK-MB cercanos o superiores al de creatinquinasa total (CK). En el presente trabajo se informa acerca de las interferencias más frecuentes encontradas con este método en nuestro Instituto
Subject(s)
Humans , Male , Female , Adult , Middle Aged , Creatine Kinase/blood , Immunologic Tests/methods , Creatine Kinase , Creatine Kinase/blood , Creatine Kinase/immunology , Creatine Kinase/isolation & purification , Electrophoresis, Cellulose Acetate/standards , Electrophoresis, Cellulose Acetate/statistics & numerical dataABSTRACT
En la práctica diaria, cuando se realiza la determinación de creatinquinasa isoenzima MB (CK-MB) por el método inmunológico, es habitual obtener resultados incongruentes: valores de CK-MB cercanos o superiores al de creatinquinasa total (CK). En el presente trabajo se informa acerca de las interferencias más frecuentes encontradas con este método en nuestro Instituto (AU)
Subject(s)
Humans , Male , Female , Adult , Middle Aged , Aged , Creatine Kinase/blood , Immunologic Tests/methods , Creatine Kinase/immunology , Electrophoresis, Cellulose Acetate/standards , Electrophoresis, Cellulose Acetate/statistics & numerical data , Creatine Kinase/isolation & purification , Creatine Kinase/blood , Creatine Kinase/diagnosisABSTRACT
Hyperinsulinemia and insulin-resistance are metabolic disturbances associated with obesity, essential hypertension, hypertriglyceridemia, glucose intolerance, overt non-insulin dependent diabetes mellitus, polymetabolic syndrome and atherosclerotic disease. The assessment of in vivo insulin sensitivity (SAI in vivo) changes achieved by life style modifications or drug interventions require a reproducible technique. To evaluate the day-to-day intra-individual repeatability of SAI-in vivo, we determined the variation in the SI index (calculated from the Minimal Model of Bergman modified by insulin or MMins) in 11 subjects with a wide range of insulin-resistance. SI (first study) varied from 0.82 to 8.48 x 10(-4) min-1/microU.mL (4.43 +/- 2.85 x 10(-4) min-1/microU.mL mean +/- SD) and highly correlated with SI (second study) (r = 0.89; p = 0.0002). The average interday coefficient of variation was 20.9 +/- 13.9% and was similar in subjects with low or high SI values. We also measured SAI in vivo by assessing the rate of serum glucose decline induced by human cristalline insulin 0.025 U/kg IV dose after a 12-14 hours fasting period (a modified Bonora's method or BBD) in 11 subjects. No subject presented biochemical or symptomatic hypoglycemia. SAI in vivo values determined by BBD varied from 21 a 234 mumol/ml/min (134 +/- 64.8 mumol/ml/min, mean +/- SD). We found a highly significant correlation between SI values obtained from MMins and SAI in vivo assessed by the BBD (r = 0.89, p = 0.0002). Our results suggest that the Mmins is a fairly reproducible procedure and that a BBD is an acceptable option to quantify SAI in vivo, mainly when a fast-execution practice is necessary or cost restrictions are required.
Subject(s)
Glucose Tolerance Test/methods , Insulin Resistance , Adult , Glucose/administration & dosage , Glucose/analysis , Humans , Hyperinsulinism , Insulin/administration & dosage , Insulin/blood , Middle Aged , Reproducibility of Results , Time FactorsABSTRACT
HTLV-I and HTLV-II are two related retroviruses that are transmitted by sexual contact, breast feeding, blood transfusion and needle sharing. In this study the prevalence of HTLV-I and HTLV-II was evaluated in voluntary blood donors as a measure of the infection in the general population. Samples were tested by a gelatine particle agglutination test and repeatedly reactive samples were confirmed by Western blot tests (WBT), enriched with recombinant rgp21, rgp461 y rgp4611 proteins, which differentiates HTLV-I and HTLV-II antibodies. Of 19,426 samples, 40 were repeatedly reactive by particle agglutination (0.21%). When analyzed by WBT, 6 met the criteria for HTLV-I (0.036%), 2 for HTLV-II (0.01%) and 1 for HTLV-I/II, 13 samples were indeterminate and 18 were negative. The prevalence is low and comparable to that from non endemic countries. Screening for anti HTLV-I/II antibodies is necessary to prevent transmission through blood transfusions.
Subject(s)
HTLV-I Infections/transmission , HTLV-II Infections/transmission , Adolescent , Adult , Aged , Agglutination Tests/methods , Argentina , Blood Donors , Blotting, Western , Female , HTLV-I Antibodies/analysis , HTLV-II Antibodies/analysis , Human T-lymphotropic virus 1/immunology , Human T-lymphotropic virus 2/immunology , Humans , Male , Middle Aged , PrevalenceABSTRACT
HTLV-I and HTLV-II are two related retroviruses that are transmitted by sexual contact, breast feeding, blood transfusion and needle sharing. In this study the prevalence of HTLV-I and HTLV-II was evaluated in voluntary blood donors as a measure of the infection in the general population. Samples were tested by a gelatine particle agglutination test and repeatedly reactive samples were confirmed by Western blot tests (WBT), enriched with recombinant rgp21, rgp461 y rgp4611 proteins, which differentiates HTLV-I and HTLV-II antibodies. Of 19,426 samples, 40 were repeatedly reactive by particle agglutination (0.21
). When analyzed by WBT, 6 met the criteria for HTLV-I (0.036
), 2 for HTLV-II (0.01
) and 1 for HTLV-I/II, 13 samples were indeterminate and 18 were negative. The prevalence is low and comparable to that from non endemic countries. Screening for anti HTLV-I/II antibodies is necessary to prevent transmission through blood transfusions.
Subject(s)
Blood Donors , HTLV-I Infections/epidemiology , HTLV-II Infections/epidemiology , Adolescent , Adult , Aged , Argentina/epidemiology , Blotting, Western , Deltaretrovirus Antibodies/blood , Female , Human T-lymphotropic virus 1/immunology , Human T-lymphotropic virus 2/immunology , Humans , Male , Mass Screening , Middle Aged , PrevalenceABSTRACT
We evaluated phagolysosomal fusion in peripheral blood monocytes from 20 HIV-infected individuals and 40 normal controls, using a fluorescence assay with acridine orange as marker. The percentages of phagolysosomal fusion of monocytes from HIV-infected subjects, after 30 and 60 min of yeast ingestion, (mean +/- standard deviation) 57.2 +/- 17 and 63.2 +/- 18.6, respectively, when compared to normal controls (72.4 +/- 7.8 and 77 +/- 8.1), did not differ significantly. However, there was a direct linear association between the percentages of phagolysosomal fusion and CD4+ lymphocytes (P < 0.001) or CD4/CD8 T-cell ratio (P < 0.01). These results suggest that phagolysosomal dysfunction becomes evident at late stages of HIV infection and progresses as CD4+.T-lymphocyte count and CD4/CD8 T-cell ratio decrease. On the other hand, recombinant gp120 inhibited significantly normal phagolysosomal fusion at concentrations ranging between 1 and 1000 ng/ml. Taking together the results obtained, we can conclude that gp120 could be responsible for monocyte phagolysosomal dysfunction observed in HIV infected patients.
