Clinica Chimica Acta最新文献_第3页

Clinical performance and application of novel serum collection “Ser-Col” device in the practice of laboratory diagnosis of infection diseases and several other immunochemical tests 新型血清采集装置 "Ser-Col "在实验室诊断感染疾病和其他几种免疫化学检验中的临床表现和应用。

IF 3.2 3区医学 Q2 MEDICAL LABORATORY TECHNOLOGY

Clinica Chimica Acta

Pub Date : 2024-09-21 DOI: 10.1016/j.cca.2024.119970

Background

Dried blood collection devices might be beneficial for diagnosing infectious diseases in areas far from the medical facilities and in lockdown situations. There are several reports on the efficacy of such applications for qualitative tests. Here we demonstrated the feasibility of a novel Ser-Col blood collection device as a standardized approach for qualitative and quantitative detection of infectious markers and several over immunochemical tests.

Methods

In the current study, we included 395 adult participants, 191 men and 204 women, with a median age of 41 years, as well as 75 children with a median age of 3 years. Serological status was determined by testing serum samples for three groups of infection diseases: hepatitis A and C, SARS-CoV-2, and herpes family viruses, as well as for thyroid peroxidase (TPO), prolactin, vitamin B12, and folate. Blood collected on the Ser-Col device (Labonovum) was eluted using an automated system (SCAUT Ser-Col automation, Blok System Supply) and manually. Ser-Col results were compared with serum sampled via standard venipuncture considered as the reference.

Results

High correlation coefficients (r = 0.95–0.99) were observed between serum samples collected with Ser-Col and via standard venipuncture for the following tests: anti-HCV, anti-SARS-CoV-2 IgG, anti-HSV-2 IgG, and anti-CMV IgM. Correlation coefficients between Ser-Col and standard venipuncture serum for anti-HSV-1 IgG, anti-CMV IgG, and anti-EBV tests were relatively low (r = 0.73–0.77). Correlation coefficients for anti-TPO, prolactin, vitamin B12, and folate were also characterized with high values (r = 0.97–0.99).

Conclusions

High accuracy and quantitative correlation were demonstrated between Ser-Col and samples collected by standard venipuncture. Hence, the Ser-Col blood collection device should be considered as a promising alternative for blood collection, storage, and transportation in both adult and pediatric populations.

背景：干式采血设备可能有利于在远离医疗设施的地区和封锁状态下诊断传染病。有几份报告指出了这种应用在定性检测方面的功效。在此，我们证明了新型血清-醇采血装置作为定性和定量检测传染病标志物以及几种免疫化学检验的标准化方法的可行性：在本次研究中，我们纳入了 395 名成年参与者，其中男性 191 人，女性 204 人，中位年龄为 41 岁；以及 75 名儿童，中位年龄为 3 岁。通过检测血清样本中的三类感染性疾病（甲型肝炎和丙型肝炎、SARS-CoV-2、疱疹病毒）、抗血小板生成素、催乳素、维生素 B12 和叶酸来确定血清学状态。使用自动系统（SCAUT Ser-Col automation，Blok System Supply）和人工洗脱在 Ser-Col 设备（Labonovum）上采集的血液。Ser-Col的结果与通过标准静脉穿刺采样的血清进行了比较：用 Ser-Col 采集的血清样本与通过标准静脉穿刺采集的血清样本在以下检测项目上的相关系数很高（r = 0.95-0.99）：抗-HCV、抗-SARS-CoV-2 IgG、抗-HSV-2 IgG 和抗-CMV IgM。在抗 HSV-1 IgG、CMV IgG 和抗 EBV 检测中，Ser-Col 与标准静脉穿刺血清之间的相关系数相对较低（r = 0.73-0.77）。抗血小板生成素、催乳素、维生素 B12 和叶酸的相关系数也具有高值（r = 0.97-0.99）的特点：Ser-Col与标准静脉穿刺采集的样本之间具有很高的准确性和定量相关性。因此，Ser-Col 采血器应被视为成人和儿童血液采集、储存和运输的理想替代品。

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引用次数: 0

Diagnostic value of six plasma biomarkers in progressive supranuclear palsy, multiple system atrophy, and Parkinson’s disease 六种血浆生物标记物对进行性核上性麻痹、多系统萎缩和帕金森病的诊断价值。

IF 3.2 3区医学 Q2 MEDICAL LABORATORY TECHNOLOGY

Clinica Chimica Acta

Pub Date : 2024-09-21 DOI: 10.1016/j.cca.2024.119975

Objectives

This study aimed to evaluate the diagnostic ability of six plasma biomarkers in progressive supranuclear palsy (PSP), multiple system atrophy (MSA), and different subtypes of Parkinson’s disease (PD).

Methods

Neurofilament light chain (NfL), phosphorylated tau-181, glial fibrillary acidic protein (GFAP), amyloid-β 42 (Aβ42), and amyloid-β 40 (Aβ40) levels were measured using the single-molecule array (Simoa) technique in a cohort of patients with PSP, MSA, different subtypes of PD, and healthy controls (HCs).

Results

Plasma NfL and GFAP levels were beneficial in discriminating between the disease groups and HCs. Plasma NfL, Aβ42, and Aβ40 could distinguish atypical Parkinsonian syndrome (APS) from PD and its subtypes. GFAP could discriminate APS from tremor dominant PD but could not discriminate APS from postural instability and gait disorder dominant PD. The efficacy of differentiation improved when a combination of multiple plasma biomarkers was applied.

Conclusions

In this study, the plasma biomarkers NfL, GFAP, Aβ42, and Aβ40 exhibited high discriminatory diagnostic value in PD and APS, and could be used as clinically potential diagnostic biomarkers. Plasma biomarker combinations could improve the differential diagnostic efficacy in the comparisons of PD and APS.

研究目的本研究旨在评估六种血浆生物标志物对进行性核上性麻痹（PSP）、多系统萎缩（MSA）和帕金森病（PD）不同亚型的诊断能力：方法：采用单分子阵列（Simoa）技术对一组帕金森病（PSP）、多系统萎缩症（MSA）、不同亚型帕金森病（PD）患者和健康对照组（HCs）的神经丝蛋白轻链（NfL）、磷酸化tau-181、胶质纤维酸性蛋白（GFAP）、淀粉样β 42（Aβ42）和淀粉样β 40（Aβ40）水平进行了测定：结果：血浆NfL和GFAP水平有利于区分疾病组和健康对照组。血浆NfL、Aβ42和Aβ40可区分非典型帕金森综合征（APS）和帕金森病及其亚型。GFAP 可将 APS 与震颤为主的帕金森病区分开来，但不能将 APS 与姿势不稳和步态障碍为主的帕金森病区分开来。当结合使用多种血浆生物标志物时，鉴别效果会更好：结论：在这项研究中，血浆生物标志物NfL、GFAP、Aβ42和Aβ40在PD和APS中表现出较高的鉴别诊断价值，可作为临床潜在的诊断生物标志物。血浆生物标记物组合可提高PD和APS比较中的鉴别诊断效果。

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引用次数: 0

Defining age-specific reference intervals for biomarkers distinguishing bacterial from viral infection in paediatrics 确定儿科细菌和病毒感染生物标志物的特定年龄参考区间。

IF 3.2 3区医学 Q2 MEDICAL LABORATORY TECHNOLOGY

Clinica Chimica Acta

Pub Date : 2024-09-21 DOI: 10.1016/j.cca.2024.119972

Differentiating bacterial from viral infections in children is a common clinical challenge. Novel host immune biomarkers have the potential to aid thediagnosis of infection aetiology and identify children who require antibiotics. Data on novel infection biomarkers gender and age-specific correlations, and reference intervals in healthy paediatrics is lacking. This study reports the plasma levels of three novel biomarkers that can aid in the differentiation of bacterial and viral infection in a healthy group of paediatrics. The levels of (Interferon-Gamma Inducible Protein 10 kDa (IP-10), Lipocalin-2 (LCN2) and TNF-Related Apoptosis-Inducing Ligand (TRAIL) were quantified in 199 plasma samples from healthy paediatrics aged 2 to 16 years old from across the UK. Reference intervals (2.5th and 97.5th) were determined, and biomarker levels were examined for sex and age associations. Reference intervals for IP-10, LCN2 and TRAIL for ages 2–16 years were 36.7–168.1 pg/ml, 14.2–123.3 ng/ml, 57.4–71.4 pg/ml respectively. No biomarker showed an association with sex and IP-10 did not show any association with age. TRAIL levels had a weak continuous negative correlation with age and LCN2 levels had a continuous positive correlation with age. Specific cut-offs for LCN2 in two age categories were identified, while TRAIL did not require age partitions. This study provides age-appropriate reference intervals for three biomarkers of infection in healthy children. These findings have the potential to improve the impact of future research on these biomarkers, the accuracy of clinical decision-making in children with infection, paediatric patient care and outcomes, and antimicrobial stewardship.

区分儿童的细菌感染和病毒感染是一项常见的临床挑战。新的免疫生物标志物有可能诊断感染病因并确定需要使用抗生素的儿童。目前还缺乏有关这些生物标志物的性别和年龄相关性以及健康儿科参考区间的数据。本研究报告了健康儿科群体中三种新型生物标记物的血浆水平，它们有助于区分细菌和病毒感染。研究人员对英国各地 199 名 2-16 岁健康儿科患者的血浆样本中干扰素-γ 诱导蛋白 10 kDa (IP-10)、脂定位蛋白-2 (LCN2) 和 TNF 相关凋亡诱导配体 (TRAIL) 的水平进行了量化。确定了参考区间（2.5th 和 97.5th），并检查了生物标记物水平与性别和年龄的关系。2-16岁儿童的IP-10、LCN2和TRAIL参考区间分别为36.7-168.1 pg/ml、14.2-123.3 ng/ml和57.4-71.4 pg/ml。没有任何生物标志物与性别相关，IP-10与年龄也没有任何关系。TRAIL水平与年龄呈弱连续负相关，LCN2水平与年龄呈连续正相关。确定了两个年龄段的 LCN2 特定临界值，而 TRAIL 则不需要年龄分区。这项研究为健康儿童的三种感染生物标志物提供了与年龄相适应的参考区间。这些发现有可能提高未来对这些生物标志物研究的影响力、感染儿童临床决策的准确性、儿科患者护理和预后以及抗菌药物管理。

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引用次数: 0

Correlation of APOE polymorphism, expression, and plasma levels with cardiac comorbidities among lipodystrophy in HIV-infected patients APOE多态性、表达和血浆水平与艾滋病病毒感染者脂肪营养不良的心脏并发症的相关性。

IF 3.2 3区医学 Q2 MEDICAL LABORATORY TECHNOLOGY

Clinica Chimica Acta

Pub Date : 2024-09-21 DOI: 10.1016/j.cca.2024.119969

Lipodystrophy in HIV-infected patients (LDHIV) includes morphological and metabolic abnormalities, including lipid and glucose metabolism. ApoE plays a role in the transport and clearance of lipoprotein. In the general population, ApoE 112 (rs429358) and 158 (rs7412) polymorphisms were linked to severe dyslipidemia. Therefore, we investigated ApoE polymorphism using PCR-RFLP in 200 HIV patients (100 with HIV-associated lipodystrophy (HIVLD), 100 without HIVLD), as well as 100 healthy controls. We also assessed ApoE expression using qRT-PCR and measured its level using ELISA. The APOE 4/4, 3/4, and 2/4 genotypes have been associated with a decreased risk of HIV-1 infection. (P = 0.0001, OR = 0.18; P = 0.006, OR = 0.87; P = 0.006, OR = 0.09) when compared between HIV-positive individuals and healthy controls. Conversely, APOE allele 2 was linked to a higher risk of acquiring HIV-1 (P = 0.03, OR = 3.02). APOE allele 2 was linked to a higher likelihood of HIVLD severity when compared between patients with and without HIVLD (P = 0.05, OR = 2.82). When comparing patients with HIVLD to healthy controls, the APOE 4/4 and 2/4 genotypes as well as allele 4 were linked with the reduced risk of LDHIV (P = 0.0006, OR = 0.21; P = 0.01, OR = 0.18; P = 0.0002, OR = 0.40). When compared to patients without HIVLD from healthy controls, the ApoE 4/4 genotype, 2 and 4 alleles, were linked to a reduced risk of developing HIVLD (P = 0.0009, OR = 0.14; P = 0.0001, OR = 0.17; P = 0.00001, OR = 0.39). When comparing impaired to normal cholesterol levels in patients without HIVLD, the ApoE 3/4 genotype was linked with the increased risk of impaired cholesterol levels (P = 0.02, OR = 3.37). When comparing impaired and normal glucose levels in patients without HIVLD, the ApoE 4/4 genotype was associated to an elevated risk of impaired glucose levels (P = 0.03, OR = 8.27). In multivariate analysis, independent impaired cholesterol, LDL, and glucose levels were associated with a higher risk of lipodystrophy severity (P = 0.04, OR = 2.33; P = 0.001, OR = 4.05; P = 0.05, OR = 2.63). ApoE expression was up-regulated in LDHIV with a fold change value of 4.02 compared to those without HIVLD. ApoE protein level was found to be higher in patients of the HIVLD group (3.01 mg/dL) compared to those without HIVLD group (2.83 mg/dL). In conclusion, individuals with ApoE allele 2 were at higher risk for HIV-1 acquisition and severity of HIVLD, whereas those with ApoE allele 4 were at reduced HIVLD severity and development risk. It’s possible that ApoE’s increased level and its overexpression are related to the ApoE allele 2 in HIVLD patients. The development of LDHIV may be facilitated by the APOE 3/4 and 4/4 genotypes as well as abnormal glucose and cholesterol levels.

艾滋病病毒感染者（LDHIV）的脂肪营养不良包括形态和代谢异常，包括脂质和葡萄糖代谢异常。载脂蛋白E在脂蛋白的转运和清除中发挥作用。在普通人群中，载脂蛋白E 112（rs429358）和158（rs7412）多态性与严重的血脂异常有关。因此，我们采用 PCR-RFLP 技术调查了 200 名艾滋病患者（其中 100 人患有艾滋病相关性脂肪变性症（HIVLD），100 人无 HIVLD）和 100 名健康对照者的载脂蛋白酶多态性。我们还使用 qRT-PCR 评估了载脂蛋白E的表达，并使用 ELISA 测定了载脂蛋白E的水平。APOE 4/4、3/4 和 2/4 基因型与 HIV-1 感染风险降低有关。(与 HIV 阳性个体和健康对照组相比，APOE 4/4、3/4 和 2/4 基因型与 HIV-1 感染风险的降低有关（P=0.0001，OR=0.18；P=0.006，OR=0.87；P=0.006，OR=0.09）。相反，APOE 等位基因 2 与感染 HIV-1 的较高风险有关（P=0.03，OR=3.02）。当将 HIVLD 患者与非 HIVLD 患者进行比较时，APOE 等位基因 2 与 HIVLD 严重程度较高的可能性有关（P=0.05，OR=2.82）。当将HIVLD患者与健康对照组进行比较时，APOE 4/4和2/4基因型以及等位基因4与LDHIV风险降低有关（P=0.0006，OR=0.21；P=0.01，OR=0.18；P=0.0002，OR=0.40）。与健康对照组中无 HIVLD 的患者相比，APOE 4/4 基因型、2 等位基因和 4 等位基因与 HIVLD 的发病风险降低有关（P=0.0009，OR=0.14；P=0.0001，OR=0.17；P=0.00001，OR=0.39）。在比较无 HIVLD 患者的胆固醇水平受损与正常时，APOE 3/4 基因型与胆固醇水平受损的风险增加有关（P=0.02，OR=3.37）。在比较无 HIVLD 患者的血糖水平受损和正常时，APOE 4/4 基因型与血糖水平受损的风险升高有关（P=0.03，OR=8.27）。在多变量分析中，独立的胆固醇、低密度脂蛋白和血糖水平受损与脂肪营养不良严重程度风险升高有关（P=0.04，OR=2.33；P=0.001，OR=4.05；P=0.05，OR=2.63）。LDHIV 中 APOE 表达上调，与未患 HIVLD 者相比，其折叠变化值为 4.02。与无 HIVLD 组（2.83 mg/dL）相比，HIVLD 组患者的 APOE 蛋白水平更高（3.01 mg/dL）。总之，具有 APOE 等位基因 2 的人感染 HIV-1 的风险较高，HIVLD 的严重程度也较高，而具有 APOE 等位基因 4 的人 HIVLD 的严重程度和发展风险较低。APOE水平的升高和过度表达可能与HIVLD患者中的APOE等位基因2有关。APOE ¾ 和 4/4 基因型以及异常的葡萄糖和胆固醇水平可能会促进 LDHIV 的发展。

{"title":"Correlation of APOE polymorphism, expression, and plasma levels with cardiac comorbidities among lipodystrophy in HIV-infected patients","authors":"","doi":"10.1016/j.cca.2024.119969","DOIUrl":"10.1016/j.cca.2024.119969","url":null,"abstract":"<div><div>Lipodystrophy in HIV-infected patients (LDHIV) includes morphological and metabolic abnormalities, including lipid and glucose metabolism. ApoE plays a role in the transport and clearance of lipoprotein. In the general population, ApoE 112 (rs429358) and 158 (rs7412) polymorphisms were linked to severe dyslipidemia. Therefore, we investigated ApoE polymorphism using PCR-RFLP in 200 HIV patients (100 with HIV-associated lipodystrophy (HIVLD), 100 without HIVLD), as well as 100 healthy controls. We also assessed <em>ApoE</em> expression using qRT-PCR and measured its level using ELISA. The <em>APOE</em> 4/4, 3/4, and 2/4 genotypes have been associated with a decreased risk of HIV-1 infection. (P = 0.0001, OR = 0.18; P = 0.006, OR = 0.87; P = 0.006, OR = 0.09) when compared between HIV-positive individuals and healthy controls. Conversely, APOE allele 2 was linked to a higher risk of acquiring HIV-1 (P = 0.03, OR = 3.02). APOE allele 2 was linked to a higher likelihood of HIVLD severity when compared between patients with and without HIVLD (P = 0.05, OR = 2.82). When comparing patients with HIVLD to healthy controls, the <em>APOE</em> 4/4 and 2/4 genotypes as well as allele 4 were linked with the reduced risk of LDHIV (P = 0.0006, OR = 0.21; P = 0.01, OR = 0.18; P = 0.0002, OR = 0.40). When compared to patients without HIVLD from healthy controls, the <em>ApoE</em> 4/4 genotype, 2 and 4 alleles, were linked to a reduced risk of developing HIVLD (P = 0.0009, OR = 0.14; P = 0.0001, OR = 0.17; P = 0.00001, OR = 0.39). When comparing impaired to normal cholesterol levels in patients without HIVLD, the <em>ApoE</em> 3/4 genotype was linked with the increased risk of impaired cholesterol levels (P = 0.02, OR = 3.37). When comparing impaired and normal glucose levels in patients without HIVLD, the <em>ApoE</em> 4/4 genotype was associated to an elevated risk of impaired glucose levels (P = 0.03, OR = 8.27). In multivariate analysis, independent impaired cholesterol, LDL, and glucose levels were associated with a higher risk of lipodystrophy severity (P = 0.04, OR = 2.33; P = 0.001, OR = 4.05; P = 0.05, OR = 2.63). <em>ApoE</em> expression was up-regulated in LDHIV with a fold change value of 4.02 compared to those without HIVLD. <em>ApoE</em> protein level was found to be higher in patients of the HIVLD group (3.01 mg/dL) compared to those without HIVLD group (2.83 mg/dL). In conclusion, individuals with <em>ApoE</em> allele 2 were at higher risk for HIV-1 acquisition and severity of HIVLD, whereas those with <em>ApoE</em> allele 4 were at reduced HIVLD severity and development risk. It’s possible that <em>ApoE</em>’s increased level and its overexpression are related to the <em>ApoE</em> allele 2 in HIVLD patients. The development of LDHIV may be facilitated by the <em>APOE</em> 3/4 and 4/4 genotypes as well as abnormal glucose and cholesterol levels.</div></div>","PeriodicalId":10205,"journal":{"name":"Clinica Chimica Acta","volume":null,"pages":null},"PeriodicalIF":3.2,"publicationDate":"2024-09-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142307237","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

A retrospective analysis of 1600 infertility patients with azoospermia and severe oligozoospermia 对1600名无精子症和严重少精子症不育患者的回顾性分析。

IF 3.2 3区医学 Q2 MEDICAL LABORATORY TECHNOLOGY

Clinica Chimica Acta

Pub Date : 2024-09-20 DOI: 10.1016/j.cca.2024.119973

Objective

This study aimed to investigate the genetic etiology of male infertility patients.

Method

A total of 1600 male patients with infertility, including 1300 cases of azoospermia and 300 cases of severe oligozoospermia, underwent routine semen analysis, chromosomal karyotype analysis and sex hormone level testing. The Azoospermia factor (AZF) on the Y chromosome was detected using the multiple fluorescence quantitative PCR technique. Additionally, copy number variation (CNV) analysis was performed on patients with Sertoli-cell-only syndrome who had a normal karyotype and AZF.

Result

Chromosomal abnormalities were found in 334 cases (20.88 %) of the 1600 male infertility patients. The most common type of abnormality was sex chromosome abnormalities (18.94 %), with 47, XXY being the most frequent abnormal karyotype. The rates of chromosomal abnormalities were significantly different between the azoospermia group and the severe oligospermia group (23.69 % and 8.67 %, respectively; P<0.05). AZF microdeletions were detected in 155 cases (9.69 %), with various deletion types and AZFc region microdeletion being the most prevalent. The rates of AZF microdeletions were not significantly different between the azoospermia group and the severe oligospermia group (9.15 % and 12 %, respectively; P=0.133). In 92 patients with Sertoli-cell-only syndrome who had a normal karyotype and AZF, the detection rate of CNV was 16.3 %. Compared to the severe oligospermia group, the azoospermia group had higher levels of FSH and LH and lower levels of T and E2, and the differences were statistically significant (P<0.05).

Conclusions

Male infertility is a complex multifactorial disease, with chromosomal abnormalities and Y chromosome microdeletions being important genetic factors leading to the disease. Initial genetic testing of infertile men should include karyotyping and Y chromosome microdeletions. If necessary, CNV testing should be performed to establish a clinical diagnosis and provide individualized treatment for male infertility.

研究目的本研究旨在探讨男性不育症患者的遗传病因：共有 1600 名男性不育症患者接受了精液常规分析、染色体核型分析和性激素水平检测，其中包括 1300 例无精子症患者和 300 例严重少精子症患者。采用多重荧光定量 PCR 技术检测了 Y 染色体上的无精子症因子（AZF）。此外，还对染色体核型和无精子症因子正常的纯绒毛膜细胞综合征患者进行了拷贝数变异（CNV）分析：结果：在 1600 名男性不育患者中，有 334 例（20.88%）发现染色体异常。最常见的异常类型是性染色体异常（18.94%），其中 47、XXY 是最常见的异常核型。无精子症组和严重少精子症组的染色体异常率有显著差异（分别为 23.69% 和 8.67%；PC 结论：男性不育是一种复杂的多因素疾病，染色体异常和Y染色体微缺失是导致该病的重要遗传因素。不育男性的初始基因检测应包括核型和 Y 染色体微缺失。如有必要，应进行 CNV 检测，以确定临床诊断，并为男性不育症提供个体化治疗。

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引用次数: 0

A stable thymidine kinase 1 tetramer for improved quality control of serum level quantification 用于改进血清水平定量质量控制的稳定胸苷激酶 1 四聚体。

IF 3.2 3区医学 Q2 MEDICAL LABORATORY TECHNOLOGY

Clinica Chimica Acta

Pub Date : 2024-09-19 DOI: 10.1016/j.cca.2024.119967

DNA synthesis is a critical process for cell growth and division. In cancer patients, an enzyme called thymidine kinase 1 (TK1) is often elevated in the blood, making it a valuable biomarker for cancer diagnosis and treatment. However, previous studies have shown that recombinant TK1 can exist in unstable mixtures of tetramers and dimers, leading to inconsistent results and potentially affecting accuracy. To address this issue, we hypothesized that incorporating tetrameric coiled-coil peptides could enhance TK1 self-assembly into stable tetramers without requiring additional adenosine triphosphate. In this study, we successfully expressed a recombinant TK1 tetramer protein in the Escherichia coli system. We optimized the induction conditions, significantly increasing protein expression levels, functionality, and solubility. Size exclusion chromatography confirmed the formation of a tetrameric structure in the expressed TK1 protein, with a molecular weight of 127.2 KDa, consistent with our expectations. We also found that the TK1 tetramer exhibited higher affinity with anti-TK1 IgY than wild-type TK1, as shown by enzyme-linked immunosorbent assay experiments. Moreover, the TK1 tetramer demonstrated good stability against heating, freeze-thawing and lyophilization with almost no immunoactivity lost. These findings suggest that recombinant TK1 tetramers have the potential to serve as calibrators in diagnostic assay kits, becoming promising candidates for quality control of clinical laboratory and in vitro diagnostic reagents.

DNA 合成是细胞生长和分裂的关键过程。在癌症患者的血液中，一种名为胸苷激酶 1（TK1）的酶通常会升高，这使其成为诊断和治疗癌症的重要生物标志物。然而，以往的研究表明，重组 TK1 可能以四聚体和二聚体的不稳定混合物形式存在，导致结果不一致，并可能影响准确性。为了解决这个问题，我们假设加入四聚体盘绕肽可以增强 TK1 自组装成稳定的四聚体，而不需要额外的三磷酸腺苷。在这项研究中，我们成功地在大肠杆菌系统中表达了重组 TK1 四聚体蛋白。我们优化了诱导条件，大大提高了蛋白质的表达水平、功能性和可溶性。尺寸排阻色谱法证实表达的 TK1 蛋白形成了四聚体结构，分子量为 127.2 KDa，与我们的预期一致。通过酶联免疫吸附实验，我们还发现 TK1 四聚体与抗 TK1 IgY 的亲和力高于野生型 TK1。此外，TK1 四聚体在加热、冻融和冻干过程中表现出良好的稳定性，几乎不丧失免疫活性。这些研究结果表明，重组 TK1 四聚体具有作为诊断试剂盒校准物的潜力，有望成为临床实验室和体外诊断试剂质量控制的候选物。

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引用次数: 0

Intra- and inter-day variations in oral metabolites from mouth-rinsed water determined using capillary electrophoresis–mass spectrometry metabolomics 利用毛细管电泳-质谱代谢组学测定漱口水中口腔代谢物的日内和日间变化。

IF 3.2 3区医学 Q2 MEDICAL LABORATORY TECHNOLOGY

Clinica Chimica Acta

Pub Date : 2024-09-14 DOI: 10.1016/j.cca.2024.119965

Background and aims

Collecting clinical samples without inconveniencing participants is desirable. The profile of metabolites in mouth-rinsed water is similar to that in saliva. However, the intra- and inter-day variations in unstimulated or stimulated saliva metabolites from mouth-rinsed water have yet to be clarified. Thus, we aimed to fill this research gap using capillary electrophoresis–mass spectrometry metabolomics.

Materials and methods

We collected mouth-rinsed water from 15 healthy participants at 9:00, 11:30, 14:00, and 16:30 daily for 3 days. In total, 509 metabolite concentrations from 180 samples were obtained using capillary electrophoresis time-of-flight mass spectrometry. Variations in each metabolite were evaluated using the Wilcoxon signed-rank test to determine at which time/day significant differences occurred after removing metabolites without significant changes using the Friedman test.

Results

Of 167 frequently detected metabolites, 100 exhibited intra-day variations, and none exhibited inter-day variations. Intra-day variations were classified into four patterns, and the intra-day variation in each metabolite was assessed. The variations may reflect elapsed time after meals, oral cleaning, or circadian rhythms.

Conclusion

This study could serve as a reference for improving the design of future clinical trials and the accuracy of metabolome analysis of mouth-rinsed water samples collected at different dates and times.

背景和目的：在不给参与者带来不便的情况下采集临床样本是可取的。漱口水中的代谢物与唾液中的代谢物相似。然而，漱口水中未受刺激或受刺激的唾液代谢物在日内和日间的变化尚未明确。因此，我们希望利用毛细管电泳-质谱代谢组学来填补这一研究空白：我们收集了 15 名健康参与者的漱口水，时间分别为每天 9:00、11:30、14:00 和 16:30，为期 3 天。使用毛细管电泳飞行时间质谱法从 180 份样本中获得了 509 种代谢物的浓度。在使用弗里德曼检验去除无显著变化的代谢物后，使用 Wilcoxon 符号秩检验对每种代谢物的变化进行评估，以确定在哪个时间/天出现显著差异：结果：在 167 种经常检测到的代谢物中，有 100 种出现了日内变化，没有一种出现日间变化。日内变化分为四种模式，并对每种代谢物的日内变化进行了评估。这些变化可能反映了餐后时间、口腔清洁或昼夜节律：本研究可作为参考，用于改进未来临床试验的设计，并提高在不同日期和时间采集的漱口水样本代谢组分析的准确性。

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引用次数: 0

Detection of egg white allergy in children by specific IgE microarray chemiluminescence immunoassay 用特异性 IgE 微阵列化学发光免疫测定法检测儿童蛋清过敏症

IF 3.2 3区医学 Q2 MEDICAL LABORATORY TECHNOLOGY

Clinica Chimica Acta

Pub Date : 2024-09-13 DOI: 10.1016/j.cca.2024.119966

Background

Allergen testing has emerged as a pivotal component in prevention and treatment strategies for allergic diseases among children and the utilization of specific IgE (sIgE) through a fully automated chemiluminescent microarray immunoassay (CLMIA) has emerged as a promising trend in the simultaneous detection of multiple allergenic components of children.

Methods

The accuracy and reliability of CLMIA were verified using children’s serum samples that concentrated on allergens. the allergens. The clinical diagnostic practicability of CLMIA was assessed through comprehensive evaluations including measurements of the limit of detection (LOD), intra-batch, and inter-batch precision, linearity analysis, the cross-contamination rate, and the concordance rate with the Phadia system.

Results

After the optimization process of CLMIA, the LODs for allergens were calculated to be below 0.01 kU/L, demonstrating the high sensitivity of CLMIA. All components exhibited good linearity within the range of 0.1–100.0 kU/L and the coefficient of determinations (R² > 0.99). The data of intra-batch precision (<10 %) and inter-batch data (<15 %) illustrated the high reproducibility of CLMIA. The cross-contamination rates for allergens (<0.5 %) showed the high accuracy of CLMIA without interfering. The positive concordance rate between CLMIA and the Phadia system exceeds 90 % with a good negative concordance rate (>85 %) and the Kappa coefficients (>0.8), suggesting the close alignment of CLMIA and the Phadia system and showing the satisfactory clinical potential of CLMIA in children’s allergy disease.

Conclusions

The application of CLMIA has been promising in allergen testing, especially for detecting multiple allergenic components in children.

背景过敏原检测已成为儿童过敏性疾病预防和治疗策略的关键组成部分，而通过全自动化学发光微阵列免疫测定（CLMIA）利用特异性 IgE（sIgE）同时检测儿童的多种过敏原成分已成为一种很有前途的趋势。通过对检测限（LOD）、批内和批间精密度、线性分析、交叉污染率以及与 Phadia 系统的吻合率进行测量等综合评价，评估了 CLMIA 的临床诊断实用性。所有成分在 0.1-100.0 kU/L 范围内线性关系良好，测定系数（R2 > 0.99）。批内精密度（10%）和批间精密度（15%）数据表明 CLMIA 具有很高的重现性。过敏原的交叉污染率（0.5%）表明 CLMIA 在无干扰的情况下具有很高的准确性。CLMIA与Phadia系统的阳性吻合率超过90%，阴性吻合率（85%）和Kappa系数（0.8）良好，表明CLMIA与Phadia系统密切吻合，显示了CLMIA在儿童过敏性疾病方面令人满意的临床潜力。

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引用次数: 0

Untargeted metabolomics analysis of the urinary metabolic signature of acute and chronic gout 对急性和慢性痛风尿液代谢特征的非靶向代谢组学分析

IF 3.2 3区医学 Q2 MEDICAL LABORATORY TECHNOLOGY

Clinica Chimica Acta

Pub Date : 2024-09-12 DOI: 10.1016/j.cca.2024.119968

Background

Gout is a common kind of inflammatory arthritis with metabolic disorders. However, the detailed pathogenesis of gout is complex and not fully clear. We investigated the urine metabolic profiling of gout patients by ultra-performance liquid chromatograph quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS).

Method

Urine metabolites were extracted from 26 acute gout patients, 31 chronic gout patients, and 32 healthy controls. Metabolite extracts were analyzed by UPLC-Q-TOF-MS for untargeted metabolomics. The peak area of creatinine was used to correct the content variations of urine samples for the semi-quantitative analysis. The value of variable importance in the projection (VIP) was obtained through the orthogonal partial least squares-discrimination analysis (OPLS-DA), and several differential metabolites were screened out.

Results

The potential metabolic markers of gout in different stages were found based on the t-test. Finally, 18 different metabolites were identified through Human Metabolome Database (HMDB) and Targeted-MS/MS. The receiver operating characteristic (ROC) curve results revealed that all the screened biomarkers exerted high accuracy and diagnostic value. Pathway analysis indicated that the significantly different metabolites were mainly involved in purine metabolism and amino acid metabolism.

Conclusion

The identified potential biomarkers are mainly involved in purine metabolism and amino acid metabolism, which leads us to further explore the pathogenesis of gout. This will lead us to further explore the pathogenesis of gout and provide the basis and ideas for the prevention and treatment of gout.

背景痛风是一种常见的伴有代谢紊乱的炎症性关节炎。然而，痛风的具体发病机制复杂且不完全清楚。我们采用超高效液相色谱四极杆飞行时间质谱（UPLC-Q-TOF-MS）对痛风患者的尿液代谢谱进行了研究。采用 UPLC-Q-TOF-MS 对代谢物提取物进行非靶向代谢组学分析。在半定量分析中，使用肌酐的峰面积来校正尿液样本的含量变化。通过正交偏最小二乘判别分析（OPLS-DA）获得了预测中的变量重要性值（VIP），并筛选出了几种差异代谢物。最后，通过人类代谢组数据库（HMDB）和靶向质谱（Targeted-MS/MS）鉴定出18种不同的代谢物。接受者操作特征曲线（ROC）结果显示，所有筛选出的生物标志物都具有很高的准确性和诊断价值。结论所发现的潜在生物标志物主要参与嘌呤代谢和氨基酸代谢，这将引导我们进一步探索痛风的发病机制。这将引导我们进一步探索痛风的发病机制，为痛风的预防和治疗提供依据和思路。

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引用次数: 0

Liquid biopsy for renal cell carcinoma 肾细胞癌的液体活检

IF 3.2 3区医学 Q2 MEDICAL LABORATORY TECHNOLOGY

Clinica Chimica Acta

Pub Date : 2024-09-10 DOI: 10.1016/j.cca.2024.119964

Liquid biopsies offer a less invasive alternative to tissue biopsies for diagnosis, prognosis, and determining therapeutic potential in renal cell carcinoma (RCC). Unfortunately, clinical studies using liquid biopsy biomarkers in RCC are limited. Accordingly, we examine RCC biomarkers, derived from urine, plasma, serum and feces of potential impact and clinical outcome in these patients. A PRISMA checklist was used to identify valuable liquid biopsy biomarkers for diagnosis (plasma cfDNA, serum- or urine-derived circulating RNAs, exosomes and proteins), prognosis (plasma cfDNA, plasma- or serum-derived RNAs, and proteins), and therapeutic response (plasma- and serum-derived proteins). Although other analytes have been identified, their application for routine clinical use remains unclear. In general, panels appear more effective than single biomarkers. Important considerations included proof of reproducibility. Unfortunately, many of the examined studies were insufficiently large and lacked multi-center rigor. Cost-effectiveness was also not available. Accordingly, it is clear that more standardized protocols need to be developed before liquid biopsies can be successfully integrated into clinical practice in RCC.

在肾细胞癌（RCC）的诊断、预后和确定治疗潜力方面，液体活检是组织活检的一种微创替代方法。遗憾的是，使用 RCC 液体活检生物标记物进行的临床研究非常有限。因此，我们研究了来自尿液、血浆、血清和粪便的 RCC 生物标记物对这些患者的潜在影响和临床结果。我们采用了 PRISMA 核对表来确定有价值的液体生物标志物，以用于诊断（血浆 cfDNA、血清或尿液衍生的循环 RNA、外泌体和蛋白质）、预后（血浆 cfDNA、血浆或血清衍生的 RNA 和蛋白质）和治疗反应（血浆和血清衍生的蛋白质）。虽然已经确定了其他分析物，但它们在常规临床中的应用仍不明确。一般来说，组合生物标志物似乎比单一生物标志物更有效。重要的考虑因素包括可重复性证明。遗憾的是，许多受检研究的规模不够大，缺乏多中心的严谨性。成本效益也无法获得。因此，在将液体活检成功纳入 RCC 临床实践之前，显然需要制定更多标准化方案。

引用次数: 0