ABSTRACT
Abstract Objective The present study aims to highlight the significance of the nucleic acid test (NAT) for musculoskeletal tissue donation and to compare the sensitivity of this test on the different available platforms. Method The present study is a retrospective survey in a human tissue bank database and an integrative literature review encompassing the last 10 years. The PubMed portal and the SCOPUS, CINAHL, and Web of Science databases were queried for articles. Results We found no specific studies on the use and sensitivity of NAT in braindead tissue donors. The information presented in the present study consists of specific contents intended for the Brazilian Blood Transfusion Network (Hemorrede Transfusional Nacional, in Portuguese) and internal retrospective data from a tissue bank located at a city in the state of São Paulo, Brazil. Conclusions The NAT is effective in blood samples from living patients. However, since biochemical reactions in braindead patients can be different, specific research, platforms, or both are crucial to tissue banks.
Resumo Objetivo Evidenciar a importância da realização do teste de ácido nucleico (NAT, na sigla em inglês) para doação de tecidos musculoesqueléticos, assim como comparar a sensibilidade deste exame nas diferentes plataformas existentes no mercado. Método Trata-se de um levantamento retrospectivo no banco de dados de um determinado Banco de Tecidos Humanos e de uma revisão integrativa da literatura, operacionalizada nos últimos 10 anos. As buscas de artigos ocorreram no portal PubMed e nas bases de dados SCOPUS, CINAHL e Web of Science. Resultados Não foram encontrados estudos específicos sobre a utilização e a sensibilidade do exame NAT em pacientes doadores de tecidos com morte encefálica (ME), sendo as informações apresentadas no presente estudo conteúdos específicos destinados à Hemorrede Transfusional Nacional e aos dados retrospectivos internos de um Banco de Tecidos do interior do estado de São Paulo, Brasil. Conclusões O exame NAT se apresenta efetivo em amostras de sangue de pacientes vivos. Porém, reações bioquímicas em pacientes com condições de ME podem se apresentar de formas diferenciadas, tornando-se indispensáveis a realização de pesquisas específicas e/ou a indicação de plataformas aos Bancos de Tecidos.
Subject(s)
Humans , Nucleic Acids , Donor SelectionABSTRACT
【Objective】 To analyze the preliminary screening and follow-up testing data of HBV in Yantai area, and discuss the rationality of following up and re-entry program of HBV reactive blood donors. 【Methods】 Donors who were single reagent reactive by enzyme-linked immunosorbent assay (ELISA) in initial screening but non-reactive by nucleic acid testing (NAT) were followed up. Individual NAT(ID-NAT) was performed for HBV DNA, ELISA for HBsAg, HBsAb, HBeAb, HBeAg and HBcAb, and ECLIA for the detection of HBsAg. 【Results】 A total of 547 blood donors were HBsAg ELISA-/NAT+, and 97 were followed up, among which 24 met the requirements of re-entry while 73 did not. Of the 24 blood donors who met the re-entry requirements, 13 donated blood again, with test results all qualified. 【Conclusion】 The combination of ELISA, ID-NAT, and ECLIA methods for following up detection for HBsAg ELISA+ blood donors is recommended. Blood donors with HbsAb S/CO ≥ 10 and negative results for other tests met the re-entry requirements, with a re-entry rate at 24.74%, and the re-donation qualified rate of blood donors after re-entry was 100%.
ABSTRACT
【Objective】 To explore the performance verification of the Shengxiang automatic NAT system for HBV DNA, HCV RNA and HIV RNA-1 using PCR-fluorescence in the laboratories of blood stations, in order to meet the requirements of T/CSBT and ensure the quality of nucleic acid detection. 【Methods】 Samples used in the external quality assessment (EQA) of National Center for Clinical Laboratories of the year 2022 were taken to verify the concordance. The standard materials of HBV DNA, HCV RNA and HIV RNA-1 were used to verify the analytical sensitivity, endogenous interfering substances, repeatability, anti-cross contamination ability and stability. 【Results】 The concordance rate of 20 EQA samples was 100%. The analytical sensitivity of HBV DNA, HCV RNA and HIV RNA-1 were all reactive and met T/CSBT. The yielding of HBV DNA, HCV RNA and HIV RNA-1 was affected little with lipemia at 3g/L and hemolysis at 4g/L. The coefficients of variation(CV) of intra-assay and inter-assay which met T/CSBT were all less than 5%, and the intra-assay variation coefficient was less than the inter-assay variation coefficient. The test results of 40 negative samples tested for cross contamination resistance were 100% negative, and 40 positive samples of HBV with 10 000 IU/mL were 100% positive. The stability verification results showed that the detection rate of weak positive samples was 100%. The coefficient of variation of the test results of the reagent after 1 and 5 freeze-thaw cycles were less than 5%,and the difference between the detection Ct value of reagent underwent once freeze-thaw and five-time freeze-thaw was not statistically significant. 【Conclusion】 The analytical sensitivity,endogenous interfering substances, repeatability,anti-cross contamination ability,stability and the compliance rate of domestic Shengxiang Gene automatic NAT system and supporting reagents by PCR-fluorescence method all meet T/CSBT, so it can be used for nucleic acid detection in blood screening in blood station laboratory.
ABSTRACT
【Objective】 To evaluate the laboratory's NAT ability by analyzing the feedback reports of nucleic acid test (NAT) results of external quality assessment (EQA) of National Center for Clinic Laboratories (NCCL), so as to improve the laboratory management details and ensure blood safety. 【Methods】 The data of NCCL NAT EQA of blood screening laboratory of Tianjin Blood Center (a total of five occasions from Jan 2019 to Jun 2021) were statistically analyzed. 【Results】 From Jan 2019 to Jun 2021, the laboratory participated in EQA for five times and all the results were qualified. The test results of NAT EQA HIV RNA/HCV RNA/HBV DNA detected by R1, R2 and R4 were consistent with the reference results. R3 showed false positive results (CT value 40.46) in the single donation detection of sample No.1925 in HCV RNA. Unreported data of the laboratory was that in the first EQA in 2021, the R4 showed false positive results (CT value 35.8) in in the single donation detection of sample No.2113 in HIV RNA. 【Conclusion】 The performance of each NAT screening system in our laboratory is relatively stable except occasional false positive results influenced by every factor. Potential problems can be found and continuously improved by assaying EQA reports and the extended experimental results of EQA samples to further improve the detection ability.
ABSTRACT
【Objective】 To explore the performance verification of NAT and its procedures for HBV DNA, HCV RNA and HIV RNA-1 using PCR-fluorescence via Cobas s201 automatic NAT system and supporting MPX V2.0 reagents that applied in the laboratories of blood stations, in order to satisfy ISO 15189 accreditation requirements and ensure the accuracy of NAT results. 【Methods】 Samples used in external quality assessment(EQA) of year 2020 were taken to verify the concordance, Performance evaluation panel and sensitivity verification panel of Roche second-generation NAT system were used to verify the sensitivity/ specificity and the lower limit of detection, respectively.And HBV DNA, HCV RNA and HIV RNA-1 quality control products were used to verify the anti-interference ability. 【Results】 The concordance rate of 40 EQA, samples was 100%. The sensitivity and specificity of Cobas s201 automatic NAT system and supporting MPX V2.0 reagents in detecting HBV DNA, HCV RNA and HIV RNA-1 were all 100%. The lower detection limit for HBV DNA, HCV RNA and HIV RNA-1 all met the requirements of reagent instructions. The yielding of HBV DNA, HCV RNA and HIV RNA-1 were affected little with hemolysis at 500 mg/dL but interfered seriously as lipemia reached 3 300 mg/dL. 【Conclusion】 The concordance rate, sensitivity, specificity and lower detection limit of the Cobas s201 fully-automatic NAT system and MPX V2.0 reagents by PCR-fluorescence method all met the requirements of reagent instructions. The verification of anti-interference ability demonstrated the requirements of ISO 15189 and the needs of blood station laboratories could be satisfied, and the detection methods and procedures can ensure the accuracy of NAT results.
ABSTRACT
【Objective】 To verify the results of HBV DNA and HCV RNA screening under different brands of vacuum collection tubes for blood samples, storage temperature and storage time. 【Methods】 Experiment 1 was conducted as follows: blood samples were collected simultaneously from 52 voluntary blood donors using two brands(divided into group A and group B) of vacuum collection tubes for blood samples. The plasma separation of group A and group B were compared, and the effects of storage time on the NAT yield of HBV DNA and HCV RNA were statistically analyzed. Experiment 2 was conducted as follows: the effects of different storage temperature, time and tubes on the NAT yield of HBV DNA and HCV RNA samples with low viral load in group A and B were verified and compared in the simulated phlebotomy condition. 【Results】 In Experiment 1: After centrifugation, blood plasma layer and cells layer were separated completely in group A(100%, 52/52), but one sample was not well separated in group B(1/52, 1.92%). After 4 to 10 h after collection, blood samples of two groups were centrifuged and screened for HBV DNA, HCV RNA within 24 h. No positive samples were yielded and the Ct values of internal control(IC-DNA and IC-RNA) were uniform. In Experiment 2: Whole blood samples, stored for either 4 h or 6~10 h at 4 ℃ or 25℃ before centrifugation, showed no difference on the NAT-yield of HBV DNA nor HCV RNA samples with low viral load(P>0.05). Ct values of HBV DNA and HCV RNA of group A was similar to those of group B as centrifuged samples were stored for 24 h or 72~104 h at 4℃(P>0.05), but all increased as the storage time prolonged. Ct values of HBV DNA in group A increased from 33.45±0.29(24 h) to 33.82±0.08(72~104 h) and HCV RNA from 35.21±0.20 to 36.12±0.43; HBV DNA from 33.46±0.25 to 34.30±0.60 and HCV RNA from 35.47±0.24 to 36.49±0.51 in group B. 【Conclusion】 Under certain laboratory condition, different storage time, storage temperature and tubes shed few effect on the NAT-yield of HBV DNA and HCV RNA samples with low virus loads. However, it is suggested that the blood sample be detected within 72 h after centrifugation at 4 ℃ storage.