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The pathogenesis and treatment strategies of acute myeloid leukemia (AML) are different for disparate gene mutations. Therefore, precise molecular testing plays a vital role in its diagnosis. However, when clinical laboratories perform molecular testing, internal quality control materials similar to clinical samples for molecular testing are lacking. At the same time, there is no related external quality assessment system to evaluate clinical laboratory test results. In order to improve the accuracy and credibility of molecular testing in clinical laboratories, we used the CRISPR/Cas9 technology to construct the DNMT3A (R882H, 2645G > A) HEK293T cell line for the quality control of AML molecular testing. We replaced the cas9 protein recognition site AGG in the ssODN with AGA to prevent the homologous recombination cell line from being cleaved by the cas9 protein again, thereby increasing the success rate of homologous recombination cell line production. It has been verified that the DNMT3A (R882H, 2645G > A) cell line can be inherited stably. The mutation frequency of the external quality assessment sample made by the DNA extraction of DNMT3A (R882H, 2645G > A) HEK293T cell line was very stable, tested by two Sanger sequencing instruments and three NGS instruments. The results above showed that the DNA extraction of DNMT3A (R882H, 2645G > A) HEK293T cell line can not only be used as internal quality control, but also be used as external quality assurance samples for monitoring different manufacturers and platforms, thereby improving the accuracy and credibility of molecular testing in clinical laboratories.
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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.
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Objective To investigate the feasibility of self‐control materials serum thyroid hormone detected in the clinical labo‐ratory internal quality control in applications .Methods The serum thyroid hormone test results in patients with high‐value ,the ser‐um was mixed with a certain proportion of the healthy population ,dubbed serum control materials ,after aliquots -20 ℃ .Every o‐riginal supporting quality control and simultaneous detection of free thyroxine (FT4) ,three free triiodothyronine (FT3) ,thyroid stimulating hormone (TSH) ,thyroglobulin (TG) ,thyroid‐microglobulin antibody (TMA) ,thyroglobulin antibodies (TGA) and other thyroid hormone six items detected before the appearance of control samples was observed continuously detected six months to observe stability ;used EXCELL do J‐L maps ,statistical mean of each project within six months ,CV and uncontrolled number of observed values in the internal quality control applications .Results Within 6 months after the product was frozen homemade quali‐ty control serum soluble appearance clear and transparent ,no precipitation and turbidity ;1 ,3 ,6 ,10 ,15 ,30 ,45 ,60 ,90 ,120 day of the first magnitude and no significant changes in thyroid hormone ;compared with the original quality control materials ,low CV values . In the six months internal quality control applications ,the cumulative results of each project without 1 ,3 ,6‐month and six months significantly different ;project and six months mean cumulative comparison between the mean drift or change in trend did not ap‐pear ;items and cumulative CV were in control of the allowable range (CV <1/3 times within total variation );6 months of internal quality control work ,runaway alarm :13s ,2 times ,22s ,1 times ,R4s ,1 times .Conclusion Serum thyroid hormone homemade frozen control samples have better stability ,and can replace its import control materials ;internal quality control in their daily activities , through EXCELL test results for J‐L diagram precision can find instability timely corrective treatment to ensure that thyroid func‐tion test results are reliable ,worthy of clinical application .
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Objective To use ELISA detecting self‐developed hepatitis B five internal quality control products .Methods Hepa‐titis B five positive sera by the detection of ELISA were diluted through optimal ratio ,and were homemade indoor quality control materials .Results Self‐control materials and commodities were simultaneously detected by ELISA ,and the test results were com‐pared ,the two were no significant difference(P>0 .05);Self‐control materials continuously detected by ELISA ,its batch variation were less than 15% ,and stability was in line with the requirements .Conclusion Self‐developed hepatitis B five indoor quality con‐trol materials are made simply ,have good stability ,are satisfied control effect ,and have promotional value .
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Objective To investigate the feasibility of self-made quality control materials on chemiluminescence analyzer .Meth-ods Residual serum in blood bags were collected and served as matrixes .Complex quality control materials of tumor markers [car-cinoembryonic antigen ,alpha-fetoprotein ,carbohydrate antigen 125(CA125) ,CA199 ,CA153 ,human chorionic gonadotrophin and total prostate specific antigen ] and hormonal markers (luteinizing hormone ,follicle-stimulating hormone ,estradiol ,testosterone , progesterone and prolactin) were prepared .The monthly average value ,coefficient of variation(CV)% value ,cumulative average value and the number of out-of-control of each marker during the 12 months were calculated .Results All self-made quality control serum which stored and packaged separately showed appearance of clear and transparent after freezing and thawing ,without turbidi-ty and sedimentation .Difference of monthly average value and cumulative average value of each marker had no statistical significance (P>0 .05) .Compared with monthly average value and cumulative average value of each marker ,no obvious drift and trends were found .Monthly indoor imprecision of each hormonal marker were controlled within the allowable range .36 times of out-of-control alarm were monitored during a year ,among them ,25 for system error and 11 for random error .Conclusion Self-made quality con-trol materials has good stability and may find the instability factors of detection system timely to guarantee the reliability of test re-sults of chemiluminescence analyzer .