Clinical application and quality management of next-generation sequencing technology / 中华检验医学杂志

With the rapid development of sequencing technology and bioinformatics analysis, the application of high-throughput sequencing (NGS) technology in precision medicine has been deepening, mainly in genetic disease diagnosis, tumor gene detection and concomitant diagnosis, as well as infectious disease detection. Compared with the traditional clinical tests, NGS analysis are more complicated with tedious operation steps and complex workflow, and they have high requirements for data analysis and variant interpretation. NGS approaches were reviewed in clinical diagnosis for constitutional disorders, cancer, infectious diseases as well as current issues in these clinical applications. Finally some suggestions for the quality management of laboratories were discussed.

Clinical application and future prospect of point-of-care testing / 中华检验医学杂志

Point-of-care testing (POCT) has the characteristics of convenient carrying, fast detection, convenient operation, high applicability to detection conditions and personnel, and can be carried out next to patients. In recent years, POCT has developed rapidly and has been widely used. The clinical value of POCT is reflected in emergency medicine, clinical front-line, primary medical care, public health prevention and control, out-of-hospital management of patients, consumer self-test and many other scenarios. With more and more extensive attention and applications, POCT is also facing challenges in its supervision and management, product quality and new scene requirements. However, with its rapid development, more advanced technologies like mobile smart devices, 5G communication technology, wearable devices, molecular and mass spectrometry detection technology, artificial intelligence are now further integrated with POCT. POCT is expected to be used for clinical diagnosis and treatment, public health prevention and control, and large-scale application. It will be widely used in health management.

Discussion on quality management of point-of-care testing / 中华检验医学杂志

Point-of-care testing (POCT) is one of the fastest growing disciplines of clinical laboratory medicine. It serves as a convenient tool for rapid clinical diagnosis. However, the increasing demand for POCT testing and the continuous expansion of the testing field has brought many new challenges to its quality management. This paper would introduce the problems existing in the clinical application of POCT, discuss the corresponding quality improvement suggestions and explore new thinkings for standardizing POCT management.

Application status and the model of quality management of point-of-care testing in Shanghai / 中华检验医学杂志

Point-of-care testing (POCT) has developed rapidly in various fields including medical health, customs quarantine, agriculture, graziery, forestry, fire-fighting, environment and food testing. It poses a huge challenge to the traditional inspection mode. Yet POCT still has many problems in terms of technical ability, quality control, regulation and industry-university-research cooperation and so on. Thus it is necessary for the health administrative departments, manufacturers and hospitals to establish consensus, and implement the total quality management system. The establishment of an independent evaluation system and the application of advanced quality control technologies of POCT is also essential given its particularity. This article focuses on the POCT application status and quality management in Shanghai based on the series of quality management measures for the POCT project carried out by the Shanghai Center for Clinical Laboratory, and raises some thoughts and suggestions for the existing problems.

Development and application of quality control materials for detection of methylated [STHX]Septin9 DNA in plasma / 临床检验杂志

Objective@#To develop a kind of quality control material which simulates clinical specimens for detecting plasma methylated Septin9 (mSEPT9) and investigate the performance for mSEPT9 detection in external quality assessment (EQA) of laboratories. @*Methods@#The cultured Hela and Jurkat cells, known to contain methylated and unmethylated Septin9 gene respectively, were cultured. The genomic DNA of the cells was collected and extracted, and detected by mSEPT9 kit. According to the Ct value, the genomic DNA was diluted into different concentrations of quality control materials with negative plasma. The homogeneity and stability of the quality control materials were evaluated. The panels consisted of 5 blindly coded samples were distributed to EQA participants and the results were summarized and evaluated. @*Results@#The purity and concentration of extracted genomic DNA met with the needs of use and could be used as quality control products for mSEPT9 detection. The homogeneity and stability met with the requirements of China National Accreditation Service for Conformity Assessment. Some of the participating laboratories occurred false positive results and false negative results, and a good linear correlation for the detected results (Ct values) was only observed in about 55.6% of the laboratories. Among the 9 participating laboratories, 7 laboratories (77.8%) performed well, 1 laboratory (11.1%) qualified, and 1 laboratory (11.1%) unqualified. @*Conclusion@#The quality control materials for mSEPT9 detection was successfully developed and the application in external quality assessment should be of great significance for evaluating and improving the detection ability of clinical laboratory.

Development and applications of quality control materials for detection of CYP3A5 genotypes / 临床检验杂志

Objective@#To evaluate the external quality assessment (EQA) program for genotyping results of tacrolimus metabolism-related cytochrome P450 family 3 subfamily A member 5 ( CYP3A5 )using plasmid DNA constructed in vitro as quality control samples, discuss the problems in clinical laboratories enrolled in the program and improve the detection quality of CYP3A5 gene. @*Methods@#Recombinant plasmid carrying CYP3A5 *3 (rs776746) AA locus sequence was constructed as wild type sample and plasmid with CYP3A5 *3 GG mutation as mutant type sample. Heterozygous mutant samples were obtained by mixing the two plasmids with equal proportion. Recombinant plasmids DNA were used as the sample panel for EQA scheme. Participating laboratories were asked to test the samples using their routine methods and report the results before deadlines. The scores of each laboratory were calculated based on their results and the overall coincidence of different samples as well as the sensitivity and specificity of different methods. @*Results@#CYP3A5 *3 locus genotypes of the constructed plasmid were verified by Sanger sequencing. The results of 15 and 17 valid laboratories were received respectively in the two EQA programs. The total percentage of 93.33% (14/15) and 100% (17/17) of the laboratories submitted correct results for all the samples. The overall coincidence rates were 96% (72/75) and 100% (85/85) respectively. All the laboratories using digital FISH got full marks in two EQA schemes, while the coincidence rates were 90% (27/30) and 100% (40/40) for Sanger sequencing. @*Conclusion@#The recombinant plasmid DNA constructed in this study could effectively detect the performance of reagents with good clinical applicability. The results of EQA programs suggested that the overall accuracy rate of enrolled laboratories was high enough, while the performances in some laboratories still need to be improved. Quality controls in clinical laboratories were essential to assure the accuracy of results.

Development and applications of external quality control materials for MTHFR 677C/T genotypes / 中华检验医学杂志

Objective To evaluate the performance of MTHFR 677 genotyping external quality assessment ( EQA) program using plasmid DNA constructed in vitro as quality control samples and discuss the problems in clinical laboratories enrolled in the program .Methods Recombinant plasmid carrying MTHFR 677C locus sequence was constructed as wild type sample and plasmid with MTHFR 677T mutation was generated with site-directed mutagenesis as mutant type sample .Heterozygous mutant samples were obtained after equal proportion of the two plasmids .EQA scheme were held twice a year in 2016 and 2017, and sample panels contained 5 different samples using recombinant plasmid DNA containing all types of MTHFR 677 locus genotypes.Participating laboratories were asked to test samples using their routine methods and report the results before deadlines .26, 28, 52 and 56 effective reports were received respectively in the four EQA schemes .The scores of each lab were calculated based on their results and the overall compliance of different samples as well as the sensitivity and specificity of different methods were calculated using Microsoft Excel .Results MTHFR 677 locus genotypes of the constructed plasmid were verified by Sanger sequencing and there was no failure of sample detection in the four EQA schemes , which suggest that the plasmid has good clinical applicability .About 96.15％( 25/26 ) ,100％( 28/28 ) ,96.15％ (50/52)and 98.21％(55/56) of the laboratories submitted correct results for all samples in the four EQA schemes.The overall compliance rate were 99.23％ ( 129/130 ) , 100％( 140/140 ) ,96.92％ ( 252/260 ) and 98.93％(277/280) respectively.All laboratories using digital FISH and microarrays got full marks in four EQA schemes.The compliance rates for fluorescent PCR were 97.5％ ( 39/40 ) , 100％ ( 45/45 ) , 94.29％(66/70) and 100％ (95/95) respectively, while the rates were 100％ (20/20), 100％ (15/15), 90％(36/40) and 92.5％(37/40) for Sanger sequencing.Conclusions The recombinant plasmid DNA constructed in this study can effectively detect the performance of reagents with good clinical applicability.The results of EQA programs suggested that the overall accuracy rate of laboratories enrolled was high enough , while some laboratories′performance still needs to be improved .Quality controls in clinical laboratories were essential to assure the accuracy of results .

Problems and reflection on quality management of clinical molecular diagnosis / 中华检验医学杂志

Gene detection related to tumor targeted therapy, pharmacogenomics, birth defects area is now widely used in clinical laboratories with the introduction of"precision medicine"concept.As next-generation-sequencing and digital PCR technology get to clinical application quickly, clinical medical decision is highly dependent on molecular diagnosis test results, therefore, the requirements for tests accuracy is unprecedented.However,the continuous development and fast expansion of molecular diagnosis technology brings outstanding quality problems.Its quality management also faces new challenges, which deserve the attention of medical laboratories and regulation bureaus.

Current mocular diagnostic technologies in drug related gene detection and its quality control system / 中华检验医学杂志

Personalized medicine has revolutionized patients ′care, and results in a personalized drug choice and dosage, through integration of pharmacology , drug information and individual ′s genetics information.The genetic testing for genes involved in drug metabolisms and targets become more and more important.However, the variety of the tests as well as the complexity of the technology brought great challenges in laboratory managements and quality assurance for individual clinical laboratory .This article briefly described the development status of pharmacogenomics and focused mainly on the routine quality assurance principle for four different techniques adopted by related genetic testing , and then presented recent research progression of related reference materials in genetic testing .Finally, the author prospects briefly the future development aspect of the quality control system .

Molecular diagnostics---the fourth revolutionary breakthrough in in vitro diagnostic technology / 中华检验医学杂志

Since the Plan of Human Genome, the project as famous asthe Manhattan Project and the Apollo Project, has finished, it brings about rapid development in molecular diagnosis and leads to revolutionary breakthrough in in-vitro diagnostics directly. Molecular diagnostics have already applicatedextensively at present in tumor , genetic diseases, congenital anomalies, hematologic diseases, infectious disease, neuropsychiatric disorders and organ transplantation .The new molecular diagnostic technologies make much contributions to clinical decision in clinic practice ,whichmeanwhile brings with new challenges to medical laboratory (clinical laboratory) as well.The medical laboratory nowadays will face the demands of high quality and expectation from clinic as well as high risks .The risk consciousness, risk management and quality control should be enhanced in medical laboratory . Strict management and standardize operations should go through pre -examination processes, examination processes and post -examination processes.Every stuff should realizes the essence of “every number is a life”.Only in this way, the medical laboratory can appropriate the development of technology and medicine and the increasing demand of clinical diagnosis , treatment, prognosis, prediction and health management.

Analysis of nonconformities in medical laboratory accreditation and status quo of clinical laboratory quality management / 中华检验医学杂志

Objective According to the analysis on nonconformities which were found from on-site assessmentin medical laboratory ISO15189accreditationcarried on byChina National Accreditation Service for Conformity Assessment(CNAS) during years 2004to 2013,to study current status of quality management in domestic medical laboratory.Methods By means of retrospective analysis, nonconformities found in 171 times of on-site audit of 133 clinical laboratories in ISO15189 accreditation during 2004 to 2013 were collected and then classified according to requirements of ISO 15189.Results Among 1 501 nonconformities involved in 171 times of on-site audit, management and technical requirements accounted for 28.5%(428) and 71.5% ( 1073 ) respectively.The mainly clauses of nonconformity were 4.3 ( 26.2%) and 4.6 (12.6%) in management requirements and 5.3 ( 25.2%), 5.4 ( 13.1%), 5.5 ( 16.0%) and 5.6 (20.2%)in technical requirements.The mainly subclauses of technical requirements were 5.3.2, 5.3.7, 5.4.3, 5.4.9, 5.5.2, 5.5.3, 5.6.1, 5.8.3 and 5.8.10.Conclusion The weakness for the medical laboratory quality management is mainly process control(5.4, 5.5 ,5.6), laboratory equipments (5.3), document control (4.3) and external services and supplies (4.6), which werethe main directions need to be improved in current medical laboratory quality management.

Research progress in international standard materials used for molecular diagnosis / 中华检验医学杂志

Benefit from the completion of human genome project and the rapid development of bioinformatics,the clinical molecular diagnostics continues to expand its scope of application which involves more widely use in infectious diseases,genetic diseases and tumors and other aspects.In the clinical molecular diagnostics,the application of international standard materials could effectively guarantee the quality of laboratory tests.This paper reviews the research progress in well-established international standard materials used for molecular diagnosis.