Chất lượng bộ sinh phẩm Polyvac - SARS-CoV-2 IgG/M ELISA chẩn đoán phát hiện kháng thể SARS - COV-2 trên các mẫu lâm sàng

2021 ◽  
Vol 31 (8) ◽  
pp. 20-29
Author(s):  
Ngô Thu Hường ◽  
Ngô Thị Thanh Hương ◽  
Trần Văn Sơn ◽  
Ngô Tiến Thọ ◽  
Vũ Thị Hường ◽  
...  
Keyword(s):  
Rt Pcr ◽  
Igm Elisa ◽  
Roche Diagnostics

Nghiên cứu này nhằm mục tiêu đánh giá  bộ sinh phẩm phát hiện kháng thể IgG/M kháng SARS-CoV-2 bằng ELISA do POLYVAC sản xuất (Polyvac SARS-CoV-2 IgG/M ELISA). Nghiên cứu được tiến hành trên 633 mẫu huyết thanh gồm 302 mẫu dương tính và 331 mẫu âm tính lấy từ bệnh nhân (2020) đã được khẳng định bằng RT-PCR. Có 200 mẫu dương tính và 200 mẫu âm tính được thực hiện so sánh với Elecsys Anti-SARS-CoV-2 của Roche Diagnostics. Kết quả cho thấy Polyvac SARS-CoV-2 IgG ELISA có độ nhạy 86,42%, độ đặc hiệu 100%; Polyvac SARS-CoV-2 IgM ELISA có độ nhạy 85,43% và độ đặc hiệu 95,77% so với RT-PCR trên các mẫu lâm sàng. Ở thời điểm sau khi có triệu chứng 0 - 7 ngày, 8 - 14 ngày, >14 ngày độ nhạy của bộ sinh phẩm tương ứng trung bình là 34,15 % ( 21,56% - 44,45%), 92,05% (84,49% - 96,17%), 97,97% (94,21% - 99,31) với IgG và 70,73%  (55,22% - 82,39%), 78,41% (68,72% - 85,72%), 92,57% (87,18% - 95,80%) với IgM. Độ tương đồng giữa Polyvac SARS-CoV-2 IgG/M ELISA và Elecsys Anti-SARS-CoV-2 với mẫu âm tính là 81% (76,49 - 86,09 %), mẫu dương tính là 98,04% (94,40 - 99,33%), độ tương đồng giữa 2 bộ sinh phẩm là 88,04% (84,45-90,83%). Polyvac SARS-CoV-2 IgG/M ELISA có thể sử dụng trong giám sát dịch tễ huyết thanh học để  xác định tiền sử có phơi nhiễm với SARS-CoV-2 và hỗ trợ chẩn đoán ở giai đoạn sau 7 ngày phát hiện có triệu chứng.

scholarly journals APPLICATION OF ELISA-IGM (MAC-ELISA) FOR DETERMINING ETIOLOGICAL LINK VIRUS TYPES IN CONCRET CASES OF THE DISEASE

2017 ◽  
Vol 22 (3) ◽  
pp. 116-121
Author(s):  
Yuliya A. Akinshina ◽  
V. F Larichev ◽  
M. A Sayfullin ◽  
S. G Mardanly ◽  
A. M Butenko
Keyword(s):  
Dengue Fever ◽  
Rt Pcr ◽  
Serum Samples ◽  
Dengue Viruses ◽  
Etiological Role ◽  
Igm Elisa ◽  
Pcr Method

105 sera from 101 patients with a confirmed diagnosis of dengue fever (LD) were examined using monospecific ELISA-IgM test. Monospecific results were observed in 39 samples (37.1%). 27 sample of them were taken during the first 7 days of the disease. In those systems were examined 23 serum samples of 23 patients, in which etiological role of one of the four dengue viruses was been established by the RT-PCR method. In this case, the coincidence-IgM ELISA results and the RT-PCR took place in 14 sera (60.9%).

scholarly journals Concomitant leptospirosis-hantavirus co-infection in acute patients hospitalized in Sri Lanka: implications for a potentially worldwide underestimated problem

2015 ◽  
Vol 143 (10) ◽  
pp. 2081-2093 ◽  
Author(s):  
N. P. SUNIL-CHANDRA ◽  
J. CLEMENT ◽  
P. MAES ◽  
H. J. DE SILVA ◽  
M. VAN ESBROECK ◽  
...  
Keyword(s):  
Retrospective Study ◽  
Sri Lanka ◽  
Clinical Outcome ◽  
Epidemiological Data ◽  
Hantaan Virus ◽  
Rt Pcr ◽  
Emerging Zoonoses ◽  
Igm Elisa ◽  
Severe Leptospirosis ◽  
Acute Patients

SUMMARYTwo global (re-)emerging zoonoses, leptospirosis and hantavirus infections, are clinically indistinguishable. Thirty-one patients, hospitalized in Sri Lanka for acute severe leptospirosis, were after exclusion of other potentially involved pathogens, prospectively screened with IgM ELISA for both pathogens. Of these, nine (29·0%) were positive for leptospirosis only, one (3·2%) for hantavirus only, seven (22·5%) for both pathogens concomitantly, whereas 13 (41·9%) remained negative for both. Moreover, in a retrospective study of 23 former patients, serologically confirmed for past leptospirosis, six (26·0%) were also positive in two different IgG ELISA hantavirus formats. Surprisingly, European Puumala hantavirus (PUUV) results were constantly higher, although statistically not significantly different, than Asian Hantaan virus (HTNV), suggesting an unexplained cross-reaction, since PUUV is considered absent throughout Asia. Moreover, RT–PCR on all hantavirus IgM ELISA positives was negative. Concomitant leptospirosis-hantavirus infections are probably heavily underestimated worldwide, compromising epidemiological data, therapeutical decisions, and clinical outcome.

scholarly journals Validation of Selected Commercial Serological Assays for Diagnosis of COVID-19

2020 ◽  
Author(s):  
Salma Younes ◽  
Hadeel Al-Jighefee ◽  
Farah Shurrab ◽  
Duaa Al-Sadeq ◽  
Hadi Yassine ◽  
...  
Keyword(s):  
Neutralization Test ◽  
Cross Reactivity ◽  
Clinical Settings ◽  
Igg Antibodies ◽  
Excellent Correlation ◽  
Rt Pcr ◽  
Rapid Assays ◽  
Igm Elisa ◽  
Rapid Tests ◽  
Antibody Tests

As researchers around the globe rush to put the available antibody tests to use, concerns have been raised about their precision. This study aimed to evaluate and compare the performance of selected commercial & automated serological assays that are widely used in different clinical settings in Qatar. We validated the performance of five commercial IgG and IgM ELISA kits, three fully automated immunoassays, and two commercial rapid tests. The sensitivity of all assays was compared to RT-PCR and a surrogate virus neutralization test (sVNT). In addition, cross-reactivity was investigated. Among the evaluated kits, Lionex IgG assay demonstrated the best performance (~88% sensitivity and ~99 specificity). All automated assays showed an excellent correlation with the neutralization test with an overall agreement of 93.6-98.5%. The rapid assays demonstrated a very good performance in detecting IgG antibodies (86.0-88.0% sensitivity and 98.0-100% specificity).

scholarly journals Antibodies to SARS-CoV2 detectable for less than 50 days in polymerase chain reaction confirmed COVID-19 patients

2020 ◽  
Vol 7 (9) ◽  
pp. 3378
Author(s):  
Nishant Kumar ◽  
Shibal Bhartiya ◽  
Tarundeep Singh
Keyword(s):  
Polymerase Chain Reaction ◽  
Healthcare Workers ◽  
Health Workers ◽  
Natural Infection ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Cross Sectional Survey ◽  
Polymerase Chain ◽  
Roche Diagnostics ◽  
Pcr Test

Background: A seroprevalence study for COVID-19 antibodies was conducted amongst health workers in Mumbai, India, in June 2020.Methods: Healthcare workers (n=801) underwent a cross sectional survey through electrochemiluminescence immunoassay (Roche diagnostics’ Elecsys anti-SARS-CoV-2 assay, Roche diagnostics, Rotkreuz, Switzerland).Results: Of the 801 healthcare workers, 62 who had been previously diagnosed with a real time-polymerase chain reaction (RT-PCR) proven SARS-CoV-2 infection, 45 (73.6%) were found to be seronegative during the study. The duration between the positive RT-PCR test and the serological testing ranged from 15 to 49 days for 34 (54.8%), and was >50 days in 28 subjects. Up to 28 days after a positive PCR test, 90% of the subjects were found to be seropositive, but this reduced to less than half over the next two weeks (38.5% between 29 and 42 days).Conclusions: Our findings are in agreement with previous reports that demonstrate a peak antibody formation after 3 weeks, and also an early antibody decay that is almost exponential. This may also have a significant effect on the protection vaccines are able to provide considering that a natural infection has such a transient antibody response. 

scholarly journals Evaluation of VIDAS® Diagnostic Assay Prototypes Detecting Dengue Virus NS1 Antigen and Anti-Dengue Virus IgM and IgG Antibodies

Diagnostics ◽  
2021 ◽  
Vol 11 (7) ◽  
pp. 1228
Author(s):  
Somphavanh Somlor ◽  
Ludovic Brossault ◽  
Marc Grandadam
Keyword(s):  
Dengue Virus ◽  
High Sensitivity ◽  
Denv Infection ◽  
Diagnostic Assay ◽  
Igg Antibodies ◽  
Rt Pcr ◽  
Igm Elisa ◽  
Ns1 Antigen ◽  
Adults And Children ◽  
Automated Platform

Dengue is a serious tropical disease caused by the mosquito-borne dengue virus (DENV). Performant, rapid, and easy-to-use assays are needed for the accurate diagnosis of acute DENV infection. We evaluated the performance of three prototype assays developed for the VIDAS® automated platform to detect dengue NS1 antigen and anti-dengue IgM and IgG antibodies. Positive and negative agreement with competitor enzyme-linked immunosorbent assays (ELISA) and rapid diagnostic tests (RDT) was evaluated in 91 Lao patients (57 adults, 34 children) with acute DENV infection. The VIDAS® NS1 assay showed the best overall agreement (95.6%) with the competitor NS1 ELISA. Both VIDAS® NS1 and NS1 ELISA assays also demonstrated high sensitivity relative to DENV RNA RT-PCR set as gold standard (85.7% and 83.9%, respectively). In contrast, NS1 RDT was less sensitive relative to DENV RNA RT-PCR (72.7%). The overall agreement of VIDAS® IgM and IgG assays with the competitor assays was moderate (72.5% for IgM ELISA, 76.9% for IgG ELISA, and 68.7% for IgM and IgG RDT). In most analyses, test agreements of the VIDAS® assays were comparable in adults and children. Altogether, the VIDAS® dengue prototypes performed very well and appear to be suitable for routine detection of dengue NS1 antigen and anti-dengue IgM/IgG antibodies.

scholarly journals Establishment of a quantitative RT-PCR detection of SARS-CoV-2 virus

2021 ◽  
Vol 26 (1) ◽  
Author(s):  
Yushen Jiang ◽  
Shanming Zhang ◽  
Hong Qin ◽  
Shuai Meng ◽  
Xuyi Deng ◽  
...  
Keyword(s):  
Limit Of Detection ◽  
Virus Detection ◽  
Quantitative Detection ◽  
Envelope Gene ◽  
Rt Pcr ◽  
Limit Of Quantification ◽  
Nucleocapsid Gene ◽  
E Gene ◽  
Standard Curve ◽  
Roche Diagnostics

Abstract Background The outbreak of novel coronavirus disease 2019 (COVID-19) has become a public health emergency of international concern. Quantitative testing of SARS-CoV-2 (severe acute respiratory syndrome coronavirus 2) virus is demanded in evaluating the efficacy of antiviral drugs and vaccines and RT-PCR can be widely deployed in the clinical assay of viral loads. Here, we developed a quantitative RT-PCR method for SARS-CoV-2 virus detection in this study. Methods RT-PCR kits targeting E (envelope) gene, N (nucleocapsid) gene and RdRP (RNA-dependent RNA polymerase) gene of SARS-CoV-2 from Roche Diagnostics were evaluated and E gene kit was employed for quantitative detection of COVID-19 virus using Cobas Z480. Viral load was calculated according to the standard curve established by series dilution of an E-gene RNA standard provided by Tib-Molbiol (a division of Roche Diagnostics). Assay performance was evaluated. Results The performance of the assay is acceptable with limit of detection (LOD) below 10E1 copies/μL and lower limit of quantification (LLOQ) as 10E2 copies/μL. Conclusion A quantitative detection of the COVID-19 virus based on RT-PCR was established.

Aurora-A Kinase Expression In Untreated Patients With Chronic Lymphocytic Leukemia

Blood ◽  
2013 ◽  
Vol 122 (21) ◽  
pp. 4896-4896
Author(s):  
Gurhan Kadikoylu ◽  
Deniz Cetin ◽  
Gokay Bozkurt ◽  
Firuzan Kacar ◽  
Irfan Yavasoglu ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Immunohistochemical Staining ◽  
Conflicts Of Interest ◽  
Deficiency Anemia ◽  
Aurora A Kinase ◽  
Aurora A ◽  
Rt Pcr ◽  
Gapdh Mrna ◽  
Roche Diagnostics ◽  
Kinase Expression

Abstract Aurora-A kinase is a cell-cyle regulating kinase required for chromosomal segregation. Overexpression of Aurora-A kinase has been detected some solid tumors and hematological malignancies such as multiple myelomai non-Hodgkin's lymhoma, and acute leukemia. But there are only two studies in chronic lymphocyic leukemia (CLL). This prospective study was approved by Ethical Committee of University. We investigated Aurora-A kinase in bone marrow of 41 untreated patients (22 male and 19 female with mean age of 70±10 years) with CLL and 19 patients (8 male and 11 female with mean age of 54±20 years) with anemia such as megaloblastic, autoimmune hemolytic, and iron deficiency anemia using a quantitative reverse transcriptase-PCR (RT-PCR) method. β-Actin and GAPDH (glyceraldehyde 3-phosphate dehydrogenase) mRNA was used as the internal controls. Total RNA was extracted from bone marrow cells using the Trizol method (High Pure Isolation Kit, Roche Diagnostics). cDNA was prepared with the Transcriptor First Strand cDNA Synthesis Kit (Roche Diagnostics). Aurora-A cDNA was quantified using TaqMan Universal PCR Mastermix (Applied Biosystems) and the Aurora-A TaqMan Gene Expression Assay. β-Actin was assayed using TaqMan Universal PCR Mastermix with forward (CCCTGGCACCCAGCAC) and reverse (GCCGATCCACACGGAGTAC) primers at 400nM each and probe (fam-ATCAAGATCATTGCTCCTCCTGAGCGC-bhq) at 100nM concentrations. Real-time quantitative RT-PCR was performed in LightCycler 480II (Roche Diagnostics). Relative RNA level was reported via standard delta delta Ct (dd Ct). Immunhistochemical analysis was performed using formalin-fixed, parafin-embedded sections of bone marrow biopsy specimens from patients and controls. Tissue sections were incubated for 60 minutes with Aurora-A (Novus Biologicals Inc. Littleton CO, 1:100 dilution). Aurora-A kinase is establish as positive if exist >10% in the cytoplasms and nuclei of the neoplastic cells in all cases. If this staining is 0-10% of cells, it is accepted as slight positive. If these cells is never (0%) stained, it is negative. For the comparison of values, Mann-Whitney-U, Chi-square, and One-Way ANOVA tests were used by SPSS 15.0 for Windows. With FISH method, 17p and 13q deletions were detected in 10% and 37% of the patients CLL, respectively. There was trisomy 12 in 7% of teh patients. 68% of the patients with CLL were in Binet-A stage. By immunohistochemical analysis, while Aurora-A kinase was positive in 61% of the patients, it was negative in 72% and slight positive 28% of controls. These positivity was statistically significant (p<0.001). By RT-PCR, β-Actin and GAPDH mRNA values were 3.85±2.61 and 3.49±2.32 in the patients with CLL, respectively. These values were 3.80±2.73 and 4.34±2.36 in controls, respectively. There was no difference for both β-Actin and GAPDH mRNA values between two groups (p>0.05). According to Binet classification, there was no difference for Aurora-A kinase expression with RT-PCR and immunohistochemical staining (p>0.05). Moreover there was no difference for both expression of Aurora-A kinase and immunhistochemical staining in between the patients with chromosmal ambnormalities and without (p>0.05). Although overexpression of Aurora-A kinase expression was not detected, significant immunohistochemical staining represented that Aurora-A kinase can be potential therapeutic target in CLL. Disclosures: No relevant conflicts of interest to declare.

scholarly journals Isolation of dengue 2 virus from a patient with central nervous system involvement (transverse myelitis)

2002 ◽  
Vol 35 (4) ◽  
pp. 401-404 ◽  
Author(s):  
Raimundo N.Q. Leão ◽  
Teichi Oikawa ◽  
Elizabeth S.T. Rosa ◽  
José T. Yamaki ◽  
Sueli G. Rodrigues ◽  
...  
Keyword(s):  
Cerebrospinal Fluid ◽  
Central Nervous System Involvement ◽  
Transverse Myelitis ◽  
Febrile Illness ◽  
Infectious Agent ◽  
Neurological Involvement ◽  
Rt Pcr ◽  
Fever Case ◽  
Igm Elisa ◽  
Acute Phase Serum

A dengue fever case is described in a 58-year-old male patient with febrile illness and thrombocytopenia complicated by neurological involvement characterized by transverse myelitis followed by weakness of both legs and flaccid paralysis. Muscle strength was much diminished and bilateral areflexia was observed. Dengue 2 (DEN-2) virus was isolated and the patient sero-converted by hemagglutination-inhibition and IgM-ELISA tests. The RT-PCR test was positive to DEN-2 in acute phase serum and culture supernatant, but negative in the cerebrospinal fluid. After three weeks of hospitalization the patient was discharged. No other infectious agent was detected in the blood and cerebrospinal fluid samples. The patient had full recovery from paralysis six months after the onset of DEN-2 infection.

scholarly journals Alta correlación en la detección de anticuerpos y antígenos de virus del dengue en muestras de suero y plasma

2015 ◽  
Vol 63 (4) ◽  
pp. 687-693
Author(s):  
Shirly Julieth Parra Alvarez ◽  
Carolina Coronel Ruiz ◽  
María Goretti Castilla Pérez ◽  
Myriam Lucia Velandia Romero ◽  
Jaime Eduardo Castellanos Parra
Keyword(s):  
Rt Pcr ◽  
Igm Elisa

<p class="p1"><span class="s1"><strong>Introducción.</strong> El diagnóstico adecuado de dengue por laboratorio es importante para la atención, así como para el control de brotes y epidemias. Hasta el momento, las pruebas ELISA para diagnóstico serológico de la infección se encuentran validadas en muestras de suero; sin embargo, en algunas ocasiones la cantidad o calidad de la muestra es inadecuada, o solo se tiene acceso a sangre anticoagulada tomada para el análisis de los parámetros hematológicos en el hemograma.</span></p><p class="p1"><strong>Objetivo.</strong> Evaluar el desempeño de cuatro pruebas ELISA en muestras de suero y plasma de casos sospechosos de dengue.  </p><p class="p1"><strong>Materiales y métodos.</strong> Se procesaron 42 muestras —21 de suero y 21 de plasma— por las técnicas de ELISA de captura para IgM, ELISA de captura para IgG, ELISA indirecta para IgG y ELISA para la detección del antígeno viral NS1. </p><p class="p1"><strong>Resultados.</strong> El porcentaje de muestras positivas encontrado fue IgM 33.3%, IgG Captura 33.3%, IgG Indirecta 90.5%, NS1 23.8%. El 42.9% de las muestras fueron positivas por RT-PCR (n=9). Todas las pruebas se comportaron igual tanto en sueros como plasmas (coeficiente Kappa 1.0). </p><p class="p1"><span class="s1"><strong>Conclusión.</strong> Los resultados obtenidos muestran una alta concordancia entre las mediciones realizadas en suero y en plasma, lo cual sugiere que la muestra de plasma puede utilizarse para el diagnóstico y la confirmación de los casos de dengue. </span></p>

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