Objective To analyze the detection ability of survival motor neuron(SMN)gene deletion of exons in clinical laboratories in Shanghai using external quality assessment(EQA).Methods Genomic DNA from the cell lines containing different SMN exon copy numbers were selected as EQA samples.The EQA sample panel,which consisted of 5 samples,was randomly selected twice and distributed to participating laboratories in Shanghai in 2023.The seresults were required to be conducted and uploaded within the specified time.Based on the reported results,statistical analysis was performed and the detection ability was evaluated.Results In the two EQA,laboratory that submitted all correct results for SMN1 accounted for 100%(38/38)and 97.22%(35/36),respectively.The overall coincidence rates were 96.92%(315/325)and 98.18%(324/330)for copy number detection of SMN1 exon 7 and 8,100%(65/65)and 96.56%(43/45)for copy number detection of SMN2 exon 7 and 8.The reported error results included case of result judgment error and 4 cases of copy number detection errors.Conclusion Screening genomic DNA from the cell lines containing different SMN exon copy numbers can effectively simulate the clinical samples,which can be applied to EQA material for deletion of exons in the SMN gene.The reported results show that the overall compliance rate of SMN deletion of exons in clinical laboratories in Shanghai is relatively high,but the testing capabilities of some laboratories still need to be improved.

Objective To evaluate the testing capabilities of laboratories,analyze existing issues,and improve testing quality,through carrying out the external quality assessment(EQA)of clinical laboratories for human papillomavirus(HPV)type 6 and 11 nucleic acid detection.Methods EQA plan was carried out twice a year.Each panel contains 4 positive samples,including one strong positive sample and one weak positive sample of HPV6 and HPV11,made from cervical secretions from patients with clinical manifestations of condyloma acuminata(CA)and positive for HPV6 or HPV11(from Shanghai First Maternity and Infant Hospital).One negative sample was cultured from the C-33A cell line(from Chinese Academy of Sciences).Samples were sent to participating laboratories by cold chain,and laboratories were required to detect test samples and upload their results within the specified time.Shanghai Center for Clinical Laboratory(SCCL)calculated the scores of each laboratory based on the return results.Results A total of 163 sample panels were sent out in the 6 rounds of EQA plan and 140 valid reports were received.The laboratory qualification rate was 96.43%(135/140)and the sample compliance rate was 97.86%(685/700).There were 13 false negative results and 2 false positive results,with weakly positive samples accounting for 76.92%(10/13)of the false negative results.Conclusion The detection accuracy of HPV6/11 nucleic acid in each laboratory was relatively high,and the detection ability of weak positive samples in individual laboratories may need to be improved.The laboratory could discover problems and improve its quality management by participating in EQA.

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.

Purpose: This study was aimed to investigate long-term survivals and toxicities of early-stage nasopharyngeal carcinoma (NPC) in endemic area, evaluating the role of chemotherapy in stage II patients. Materials and Methods: Totally 187 patients with newly diagnosed NPC and restaged American Joint Committee on Cancer/ International Union Against Cancer 8th T1-2N0-1M0 were retrospectively recruited. All received intensity-modulated radiotherapy (IMRT)±chemotherapy (CT) from 2001 to 2010. Results: With 15.7-year median follow-up, 10-year locoregional recurrence-free survival, distant metastasis-free survival (DMFS), disease-specific survival (DSS), and overall survival (OS) were 93.3%, 93.5%, 92.9% and 88.2%, respectively. Multivariable analyses showed cervical lymph nodes positive and pre-treatment prognostic nutritional index ≥ 52.0 could independently predict DMFS (p=0.036 and p=0.011), DSS (p=0.014 and p=0.026), and OS (p=0.002 and p < 0.001); Charlson comorbidity index < 3 points could predict DSS (p=0.011); age > 45 years (p=0.002) and pre-treatment lactate dehydrogenase ≥ 240 U/L (p < 0.001) predicted OS. No grade 4 late toxicity happened; grade 3 late toxicities included subcutaneous fibrosis (4.3%), deafness or otitis (4.8%), skin dystrophy (2.1%), and xerostomia (1.1%). No differences on survivals were shown between IMRT+CT vs. IMRT alone in stage II patients, even in T2N1M0 (p > 0.05). Unsurprising, patients in IMRT+CT had more acute gastrointestinal reaction, myelosuppression, mucositis, late ear toxicity, and cranial nerve injury (all p < 0.05) than IMRT alone group. Conclusion Superior tumor control and satisfying long-term outcomes could be achieved with IMRT in early-stage NPC with mild late toxicities. As CT would bring more toxicities, it should be carefully performed to stage II patients.

NKX6-2 encodes a transcription factor involving the genesis and development of oligodendrocytes.Mutations in the NKX6-2 gene are related to a severe variant of hypomyelinating leukodystrophy, namely the NKX6-2-related spastic ataxia type 8 (SPAX8). As of March 24, 2020, there are 36 cases of SPAX8 involving 11 mutations of the NKX6-2 gene, which has not been reported in Chinese population.This article for the first time reported a 31-month-old Chinese boy diagnosed as SPAX8 caused by a novel mutation of the NKX6-2 gene at c. 234dup(p.Leu79Alafs*? ) admitted to the Department of Rehabilitation, Guangdong Women and Children Hospital in October 2019 retrospectively.He presented severe global psychomotor delay, spasticity, nystagmus, hearing impairment and trichiasis, ametropia of both eyes, and retinal degeneration of the right eye, which have not been reported in previous cases.His elder sister presented relevant manifestations, but did not perform the genetic testing.Through this case report and literature review, the genotype and phenotype of SPAX8 were expanded.

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.

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.

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.

A male patient, with diagnose of Madelung disease, was admitted in September 2009. He has been a heavy drinker for decades before onset of the disease. This patient was characterized by the large amount of symmetrical deposits of adipose tissue in the subcutaneous layer around neck, and without obesity on other sites. The excessive adipose tissue was surgically removed by three steps. Appearance almost returned to normal. No recurrence happened after 8 years of follow-up.

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.