Practical Laboratory Medicine最新文献

英文中文

Integrative analysis of exosomal ncRNAs and their regulatory networks in liver cancer progression

IF 1.7 Q3 MEDICAL LABORATORY TECHNOLOGY

Practical Laboratory Medicine

Pub Date : 2025-03-21 DOI: 10.1016/j.plabm.2025.e00464

Farzin Mirzaei-nasab , Ahmad Majd , Yousef Seyedena , Nazanin Hosseinkhan , Najma Farahani , Mehrdad Hashemi

Background

Hepatocellular carcinoma (HCC) is a significant global health challenge with complex molecular underpinnings. Recent advancements in understanding the role of non-coding RNAs (ncRNAs) and exosomes in cancer biology have opened new avenues for research into potential diagnostic and therapeutic strategies.

Methods

This study utilized a comprehensive approach to analyze gene expression patterns and regulatory networks in HCC. We integrated RNA sequencing data gathered from both tissue samples and exosomes. The WGCNA and limma R packages were employed to construct co-expression networks and identify differentially expressed ncRNAs, including long non-coding RNAs (lncRNAs) and circular RNAs (circRNAs).

Results

Our analysis demonstrated distinct expression profiles of various ncRNAs in HCC, revealing their intricate interactions with cancer-related genes. Key findings include the identification of a network of microRNAs that interact with selected lncRNAs and their potential roles as biomarkers. Moreover, exosomal RNA was shown to effectively reflect tissue-specific gene expression changes.

Conclusions

The results of this study highlight the significance of exosomal ncRNAs in the progression of liver cancer, suggesting their potential as both diagnostic biomarkers and therapeutic targets. Future research should focus on the functional implications of these ncRNAs to further elucidate their roles in HCC and explore their applications in clinical settings.

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引用次数: 0

A 9-year-old child presenting with anemia accompanied by abnormal red blood cell morphology

IF 1.7 Q3 MEDICAL LABORATORY TECHNOLOGY

Practical Laboratory Medicine

Pub Date : 2025-03-20 DOI: 10.1016/j.plabm.2025.e00459

Huijun Qin , Yuan He , Zaixiang Xie

Cases of anemia presenting with abnormal erythrocyte morphology often pose diagnostic challenges, particularly in patients with refractory anemia. Here, we present the case of a 9-year-old male patient under investigation for anemia, who had a history of anemia and received a blood transfusion at birth. Despite the absence of obvious clinical manifestations related to anemia thereafter, his condition was not given due consideration. The patient experienced a sudden onset of illness and was initially suspected to have thalassemia. However, subsequent pertinent examinations, notably bone marrow aspiration and genetic testing, led to the diagnosis of hereditary sideroblastic anemia alongside chronic atrophic gastritis. This case illustrates the diagnostic journey of anemia characterized by abnormal red blood cell morphology, aiming to facilitate early and accurate diagnosis, as well as prompt treatment, for such patients in clinical practice.

引用次数: 0

Evaluation of CircHIPK3 biomarker potential in breast cancer

IF 1.7 Q3 MEDICAL LABORATORY TECHNOLOGY

Practical Laboratory Medicine

Pub Date : 2025-03-20 DOI: 10.1016/j.plabm.2025.e00470

Ensiyeh Bahadoran , Davood Mohammadi , Manijeh Jalilvand , Sahar Moghbelinejad

Background

Nowadays, the investigation of circular RNAs (circRNAs) in various cancers is of great interest. In this research, we evaluated circHIPK3 biomarker potential in breast cancer (BC).

Methods

The studied samples were 100 cancer and adjacent normal tissues, plasma from 95 cancer patients, 42 patients with fibroadenomatosis, and 93 healthy donors. Illumina high-throughput Hi Seq 2000 sequencing performed expression profiling on 4 pairs of cancerous and normal breast tissues. For expression confirmation, Quantitative real-time fluorescent polymerase chain reaction (qRT-PCR) was used to detect the expression level of circHIPK3. CircHIPK3 diagnostic efficacy was evaluated by the receiver operating characteristic curve (ROC).

Results

Based on high-throughput sequencing and bioinformatics results circHIPK3 had the highest expression in cancer tissues (P = 0.00034). Real-time results showed expression upregulation of circHIPK3 in BC tissues and plasma in comparison to healthy controls (P < 0.0001). For diagnostic potential, the area under the curve (AUC) result was 0.8087 (95 % CI: 0.7309 to 0.8866, P < 0.0001). Also, our results showed high specificity and sensitivity of circHIPK3 when evaluated alongside the CA-15-3 and CEA. Pathologic criteria evaluation showed that upregulation of circHIPK3 correlates with tumor size.

Conclusions

CircHIPK3 is significantly upregulated in BC tissues and plasma compared to healthy controls, demonstrating high diagnostic potential with an AUC of 0.8087. The expression of circHIPK3 correlates with tumor size, indicating its relevance in the pathologic assessment of BC.

{"title":"Evaluation of CircHIPK3 biomarker potential in breast cancer","authors":"Ensiyeh Bahadoran , Davood Mohammadi , Manijeh Jalilvand , Sahar Moghbelinejad","doi":"10.1016/j.plabm.2025.e00470","DOIUrl":"10.1016/j.plabm.2025.e00470","url":null,"abstract":"<div><h3>Background</h3><div>Nowadays, the investigation of circular RNAs (circRNAs) in various cancers is of great interest. In this research, we evaluated circHIPK3 biomarker potential in breast cancer (BC).</div></div><div><h3>Methods</h3><div>The studied samples were 100 cancer and adjacent normal tissues, plasma from 95 cancer patients, 42 patients with fibroadenomatosis, and 93 healthy donors. Illumina high-throughput Hi Seq 2000 sequencing performed expression profiling on 4 pairs of cancerous and normal breast tissues. For expression confirmation, Quantitative real-time fluorescent polymerase chain reaction (qRT-PCR) was used to detect the expression level of circHIPK3. CircHIPK3 diagnostic efficacy was evaluated by the receiver operating characteristic curve (ROC).</div></div><div><h3>Results</h3><div>Based on high-throughput sequencing and bioinformatics results circHIPK3 had the highest expression in cancer tissues (<em>P</em> = 0.00034). Real-time results showed expression upregulation of circHIPK3 in BC tissues and plasma in comparison to healthy controls (<em>P</em> < 0.0001). For diagnostic potential, the area under the curve (AUC) result was 0.8087 (95 % CI: 0.7309 to 0.8866, <em>P</em> < 0.0001). Also, our results showed high specificity and sensitivity of circHIPK3 when evaluated alongside the CA-15-3 and CEA. Pathologic criteria evaluation showed that upregulation of circHIPK3 correlates with tumor size.</div></div><div><h3>Conclusions</h3><div>CircHIPK3 is significantly upregulated in BC tissues and plasma compared to healthy controls, demonstrating high diagnostic potential with an AUC of 0.8087. The expression of circHIPK3 correlates with tumor size, indicating its relevance in the pathologic assessment of BC.</div></div>","PeriodicalId":20421,"journal":{"name":"Practical Laboratory Medicine","volume":"45 ","pages":"Article e00470"},"PeriodicalIF":1.7,"publicationDate":"2025-03-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143682981","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Gene variant analysis in pediatrics with early-onset epilepsy: Identification of novel variants

IF 1.7 Q3 MEDICAL LABORATORY TECHNOLOGY

Practical Laboratory Medicine

Pub Date : 2025-03-19 DOI: 10.1016/j.plabm.2025.e00462

Pooyan Alizadeh , Armin Jahangiri Babadi , Nemat Ghadiri , Mostafa Neissi , Masoud Zeinali

Background

Epilepsy encompasses a range of brain disorders, often accompanied by growth delay and cerebral palsy. The identification of gene variants is critical for guiding treatment strategies in patients with epilepsy. This study investigates the genetic variants in patients with early-onset epilepsy (EOE) through whole-exome sequencing (WES).

Materials and methods

DNA was extracted from peripheral blood using a standard salting-out method. Gene variants were identified using WES, and sequencing data were analyzed through a two-step approach.

Results

Among 20 subjects, WES identified two novel variants. The first variant, AP3B2 (NM_001278512.2: c.3190G > A; p. Val1064Ile), was located in exon 27 and exhibited homozygosity in the proband and heterozygosity in the parents. The second variant, PIGB (NM_004855.5: c.1664G > C; p.Ter555Serext∗54), was located in exon 12 and demonstrated a similar inheritance pattern. Notably, the PIGB variant was associated with elevated ALP levels.

Conclusion

This study highlights the value of WES in identifying genetic variants associated with epilepsy, particularly the novel AP3B2 and PIGB variants. By focusing on these impactful findings, the study advances understanding of epilepsy genetics and emphasizes the role of WES in enabling early diagnosis, personalized treatment, and improved management strategies.

{"title":"Gene variant analysis in pediatrics with early-onset epilepsy: Identification of novel variants","authors":"Pooyan Alizadeh , Armin Jahangiri Babadi , Nemat Ghadiri , Mostafa Neissi , Masoud Zeinali","doi":"10.1016/j.plabm.2025.e00462","DOIUrl":"10.1016/j.plabm.2025.e00462","url":null,"abstract":"<div><h3>Background</h3><div>Epilepsy encompasses a range of brain disorders, often accompanied by growth delay and cerebral palsy. The identification of gene variants is critical for guiding treatment strategies in patients with epilepsy. This study investigates the genetic variants in patients with early-onset epilepsy (EOE) through whole-exome sequencing (WES).</div></div><div><h3>Materials and methods</h3><div>DNA was extracted from peripheral blood using a standard salting-out method. Gene variants were identified using WES, and sequencing data were analyzed through a two-step approach.</div></div><div><h3>Results</h3><div>Among 20 subjects, WES identified two novel variants. The first variant, <strong>AP3B2</strong> (NM_001278512.2: c.3190G > A; p. Val1064Ile), was located in exon 27 and exhibited homozygosity in the proband and heterozygosity in the parents. The second variant, <strong>PIGB</strong> (NM_004855.5: c.1664G > C; p.Ter555Serext∗54), was located in exon 12 and demonstrated a similar inheritance pattern. Notably, the <strong>PIGB</strong> variant was associated with elevated ALP levels.</div></div><div><h3>Conclusion</h3><div>This study highlights the value of WES in identifying genetic variants associated with epilepsy, particularly the novel AP3B2 and PIGB variants. By focusing on these impactful findings, the study advances understanding of epilepsy genetics and emphasizes the role of WES in enabling early diagnosis, personalized treatment, and improved management strategies.</div></div>","PeriodicalId":20421,"journal":{"name":"Practical Laboratory Medicine","volume":"45 ","pages":"Article e00462"},"PeriodicalIF":1.7,"publicationDate":"2025-03-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143734855","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Technical improvement on next-generation sequencing in clinical application

IF 1.7 Q3 MEDICAL LABORATORY TECHNOLOGY

Practical Laboratory Medicine

Pub Date : 2025-03-08 DOI: 10.1016/j.plabm.2025.e00465

Xiaolei Xie, Weiguo Yin, Shuxia Xuan, Fuguang Li

The next-generation sequencing (NGS) technology is currently widely utilized in clinical laboratories. This paper describes a method of technical improvement on NGS, which increases the success rates of NGS detection. This study reveals that the DNA library concentration is the highest at a molar ratio of 100:1 for the adapter to DNA. The self-revised method for adaptor ligation can effectively improve the success rate of DNA library, particularly in manual operations. Additionally, the modified pooling method, which incorporates various DNA fragment sizes for different NGS projects, proves beneficial for medical laboratory applications.

引用次数: 0

Indirect reference intervals for TSH in a sample of lebanese pregnant women

IF 1.7 Q3 MEDICAL LABORATORY TECHNOLOGY

Practical Laboratory Medicine

Pub Date : 2025-03-08 DOI: 10.1016/j.plabm.2025.e00460

Dollen Eid , Nizar El Bcherawi , Georges Abi Tayeh , Nada El Ghorayeb , Marie-Hélène Gannagé-Yared

Background

Thyroid dysfunction in pregnant women can lead to fetal complications. Thyroid-stimulating hormone (TSH) is the key hormone for diagnosis of thyroid dysfunction. No previous study has established reference intervals (RI) for TSH in Lebanese pregnant women. The objective of this study is to define the TSH RIs for each trimester of pregnancy in healthy Lebanese pregnant women using an indirect method.

Materials and methods

This retrospective study included 287 pregnancies selected from the records of an obstetric clinic at Hôtel-Dieu de France University Hospital from January 2021 to May 2023. A control group of 103 non-pregnant women was also included in the study. The collected TSH values were stratified by trimester (first and second) of pregnancy and postpartum. After applying the exclusion criteria, a total of 458 TSH values were included in the analysis.

Results

The respective medians and RIs for TSH during the first, second pregnancy trimesters and postpartum are 1.57 (0.43–3.20 mIU/L), 1.84 (0.56–4.41 mIU/L), and 1.38 (0.30–3.60 mIU/L), while for the control group it is 1.66 (0.64–4.24 mIU/L). There is a significant correlation between TSH values in the first trimester and those in the second trimester and postpartum (p ≤ 0.001 and p = 0.002 respectively). No significant correlation was observed between age and TSH levels in the first and second trimesters and as well as in postpartum.

Conclusion

Our RIs are close to the revised American Thyroid Association (ATA) recommendations. Further research is needed to understand the mechanisms and clinical impact of these differences.

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引用次数: 0

The value of loop-mediated isothermal amplification in diagnosing lower respiratory tract infections in children

IF 1.7 Q3 MEDICAL LABORATORY TECHNOLOGY

Practical Laboratory Medicine

Pub Date : 2025-03-08 DOI: 10.1016/j.plabm.2025.e00463

Feng Yan, Shikun Xu, Meijing Shen, Yu Zhao, Huabo Tong, Kaifeng Wu, He Zha

Objective

To evaluate the diagnostic value of loop-mediated isothermal amplification(LAMP) chip method (hereinafter referred to as "LAMP") in the detection of pathogens in children with lower respiratory tract infections(LRTIs).

Methods

Sputum samples from 1723 children with LRTIs hospitalized from April 2020 to April 2021 were collected. Pathogen detection was performed using both LAMP and sputum culture method(SCM).Detection rates and consistency between the two methods were analyzed using the Chi-square test and Kappa analysis.

Results

The positive detection rates of the LAMP and the SCM were 58.97 %(1016/1723) and 43.64 %(752/1723), respectively(P<0.001). The detection rates of Streptococcus pneumoniae (Spn)(24.26 %/13.52 %), Staphylococcus aureus(Sau)(13.12 %/10.39 %), Acinetobacter baumannii (Aba)(1.33 %/0.48 %), Stenotrophomonas maltophilia (Sma)(0.58 %/0.12 %), and Haemophilus influenzae(Hin)(31.05 %/16.19 %) were significantly higher with the LAMP than with the SCM(P<0.05). Both methods showed that single infections were predominant among children, with positive rates of 65.06 % and 87.23 %, respectively, with Hin(49.92 %/33.69 %) being the most common pathogen.In mixed infections, the positive rates were 34.94 % and 12.77 %, respectively, with mixed infections of Hin and Spn being the most common, accounting for 48.89 % and 32.29 % of cases, respectively. There were significant differences in the detection rates of Spn, Sau, Klebsiella pneumoniae(Kpn), Sma, Hin, and Escherichia coli(Eco) between single and mixed infections(P < 0.05). The detection results of Spn, Sau, Kpn, Hin, and Eco exhibited high consistency between the two methods, while the consistency for Pseudomonas aeruginosa(Pae), Aba, and Sma was lower.

Conclusion

The LAMP is simpler, faster, more sensitive and specific than SCM, offering a reliable laboratory diagnostic basis for clinical management of LRTIs in children.

目的评价环介导等温扩增芯片法（LAMP）（以下简称 "LAMP"）在下呼吸道感染（LRTI）患儿病原体检测中的诊断价值。方法收集2020年4月至2021年4月期间住院的1723名LRTI患儿的痰标本。结果 LAMP 和 SCM 的阳性检出率分别为 58.97 %（1016/1723）和 43.64 %（752/1723）（P<0.001）。肺炎链球菌（Spn）（24.26 %/13.52 %）、金黄色葡萄球菌（Sau）（13.12 %/10.39 %）、鲍曼不动杆菌（Aba）（1.33 %/0.48 %）、嗜麦芽血单胞菌（Sma）（0.58 %/0.12 %）和流感嗜血杆菌（Hin）（31.05 %/16.19 %）。两种方法均显示，单一感染在儿童中占主导地位，阳性率分别为 65.06 % 和 87.23 %，其中 Hin（49.92 %/33.69 %）是最常见的病原体；在混合感染中，阳性率分别为 34.94 % 和 12.77 %，其中 Hin 和 Spn 混合感染最常见，分别占 48.89 % 和 32.29 %。Spn、Sau、肺炎克雷伯菌（Kpn）、Sma、Hin和大肠埃希菌（Eco）的检出率在单一感染和混合感染之间存在明显差异（P <0.05）。两种方法对 Spn、Sau、Kpn、Hin 和 Eco 的检测结果一致性较高，而对铜绿假单胞菌（Pae）、Aba 和 Sma 的检测结果一致性较低。

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引用次数: 0

Biogenic amine testing in the South African public health care system

IF 1.7 Q3 MEDICAL LABORATORY TECHNOLOGY

Practical Laboratory Medicine

Pub Date : 2025-01-28 DOI: 10.1016/j.plabm.2025.e00457

D. Legg-E’Silva , E.M. Cave , T. Snyman, S. Currin, N. Kone, K.L. Prigge

Background

Phaeochromocytoma, paraganglioma and neuroblastoma are catecholamine secreting neuroendocrine tumours. Biochemical screening for suspected cases of these tumours involves the measurement of catecholamines and their metabolites in either urine or plasma. The South African National Health Laboratory service (NHLS) measures urine fractionated metanephrines (UMF) and normetanephrines (UNF), urine vanillylmandelic acid (UVMA) and urine homovanillic acid (UHVA).

Objectives

To analyse the demographic, biochemical and testing patterns of patients’ UMF, UNF, UVMA and UHVA in the NHLS.

Methods

Data from January 2015 to December 2016 for all patients undergoing UMF, UNF, UVMA and UHVA testing was extracted from the NHLS central data warehouse. Neuroendocrine tumours were biochemically diagnosed when results were >2x multiples of the upper reference limits. Multiple testing was defined as ≥2 tests within a 14-day period. Ethnicity was determined through hot-deck imputation.

Results

Biochemically abnormal test results were identified by UMF/UNF measurements in 98.2 % of cases. In 1.8 % of cases, the addition of UVMA resulted in a previously unidentified biochemical positive. Adult white and coloured populations have significantly less biochemically positive UMF results compared to the African population. Multiple testing resulted in discordant results for 12.8 % of UMF and 13.1 % of UNF testing.

Conclusion

UVMA testing for phaeochromocytoma and paraganglioma offers little benefit over testing with UMF alone. Requesting consecutive multiple samples is preferred, however, a single 24-h fractionated UMF/UNF is efficient and cost-effective for phaeochromocytoma and paraganglioma screening, with further testing recommended when clinically indicated. African individuals are more likely to have raised catecholamines and requires further investigation.

{"title":"Biogenic amine testing in the South African public health care system","authors":"D. Legg-E’Silva , E.M. Cave , T. Snyman, S. Currin, N. Kone, K.L. Prigge","doi":"10.1016/j.plabm.2025.e00457","DOIUrl":"10.1016/j.plabm.2025.e00457","url":null,"abstract":"<div><h3>Background</h3><div>Phaeochromocytoma, paraganglioma and neuroblastoma are catecholamine secreting neuroendocrine tumours. Biochemical screening for suspected cases of these tumours involves the measurement of catecholamines and their metabolites in either urine or plasma. The South African National Health Laboratory service (NHLS) measures urine fractionated metanephrines (UMF) and normetanephrines (UNF), urine vanillylmandelic acid (UVMA) and urine homovanillic acid (UHVA).</div></div><div><h3>Objectives</h3><div>To analyse the demographic, biochemical and testing patterns of patients’ UMF, UNF, UVMA and UHVA in the NHLS.</div></div><div><h3>Methods</h3><div>Data from January 2015 to December 2016 for all patients undergoing UMF, UNF, UVMA and UHVA testing was extracted from the NHLS central data warehouse. Neuroendocrine tumours were biochemically diagnosed when results were >2x multiples of the upper reference limits. Multiple testing was defined as ≥2 tests within a 14-day period. Ethnicity was determined through hot-deck imputation.</div></div><div><h3>Results</h3><div>Biochemically abnormal test results were identified by UMF/UNF measurements in 98.2 % of cases. In 1.8 % of cases, the addition of UVMA resulted in a previously unidentified biochemical positive. Adult white and coloured populations have significantly less biochemically positive UMF results compared to the African population. Multiple testing resulted in discordant results for 12.8 % of UMF and 13.1 % of UNF testing.</div></div><div><h3>Conclusion</h3><div>UVMA testing for phaeochromocytoma and paraganglioma offers little benefit over testing with UMF alone. Requesting consecutive multiple samples is preferred, however, a single 24-h fractionated UMF/UNF is efficient and cost-effective for phaeochromocytoma and paraganglioma screening, with further testing recommended when clinically indicated. African individuals are more likely to have raised catecholamines and requires further investigation.</div></div>","PeriodicalId":20421,"journal":{"name":"Practical Laboratory Medicine","volume":"44 ","pages":"Article e00457"},"PeriodicalIF":1.7,"publicationDate":"2025-01-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143140812","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Preparation of recombinant myoglobin and investigation of the liquid antigen stability for quality control materials

IF 1.7 Q3 MEDICAL LABORATORY TECHNOLOGY

Practical Laboratory Medicine

Pub Date : 2025-01-28 DOI: 10.1016/j.plabm.2025.e00456

Yu-Hui Wang, Xi-Feng Sun, Chun-Xin Xu, Feng-Qiang Sun, Rong-Rong Wang, Xiao-Kun Bian, Zhan-Zhao Wang, Qiang Wu

Myoglobin (Mb) has been used as a biomarker for acute myocardial infarction. This study aimed to evaluate the stability of liquid Mb as quality control materials for Mb determination. Mb protein was expressed in Escherichia coli system and purified using Ni²⁺ chelate affinity chromatography. The purity of purified recombinant Mb reached to 95 %. The immunoreactivity of Mb was investigated using Mb assay kits. The coefficient of determination (R²) of the curve fitted with dilution ratio and Mb concentration as variables was greater than 0.95, which indicated that Mb had good immunoreactivity. The concentrations per gradient measured using different kits had no significant difference (p-value>0.05), which indicated that the reactivity between the Mb antigen and Mb antibodies with different epitopes was good. The effects of different storage buffer, storage temperature and storage times on the stability of liquid Mb were investigated by detecting the concentration changes. At 2–8 °C for two months, Mb concentration in buffer B (Tris-HCl, pH 7.8, containing 1 % BSA and 0.05 % NaN₃) decreased within 10 % compared with the initial concentration. The long-term storage stability was investigated by the thermal acceleration experiment. At 37 °C for one week, Mb concentration decreased by less than 15 %, indicating that the Mb had good long-term storage stability. The prepared liquid Mb had good immunoreactivity and stability, avoiding storage in freeze-dried powder. It was a promising alternative as the quality control material for Mb detection.

{"title":"Preparation of recombinant myoglobin and investigation of the liquid antigen stability for quality control materials","authors":"Yu-Hui Wang, Xi-Feng Sun, Chun-Xin Xu, Feng-Qiang Sun, Rong-Rong Wang, Xiao-Kun Bian, Zhan-Zhao Wang, Qiang Wu","doi":"10.1016/j.plabm.2025.e00456","DOIUrl":"10.1016/j.plabm.2025.e00456","url":null,"abstract":"<div><div>Myoglobin (Mb) has been used as a biomarker for acute myocardial infarction. This study aimed to evaluate the stability of liquid Mb as quality control materials for Mb determination. Mb protein was expressed in <em>Escherichia coli</em> system and purified using Ni<sup>2+</sup> chelate affinity chromatography. The purity of purified recombinant Mb reached to 95 %. The immunoreactivity of Mb was investigated using Mb assay kits. The coefficient of determination (R<sup>2</sup>) of the curve fitted with dilution ratio and Mb concentration as variables was greater than 0.95, which indicated that Mb had good immunoreactivity. The concentrations per gradient measured using different kits had no significant difference (p-value>0.05), which indicated that the reactivity between the Mb antigen and Mb antibodies with different epitopes was good. The effects of different storage buffer, storage temperature and storage times on the stability of liquid Mb were investigated by detecting the concentration changes. At 2–8 °C for two months, Mb concentration in buffer B (Tris-HCl, pH 7.8, containing 1 % BSA and 0.05 % NaN<sub>3</sub>) decreased within 10 % compared with the initial concentration. The long-term storage stability was investigated by the thermal acceleration experiment. At 37 °C for one week, Mb concentration decreased by less than 15 %, indicating that the Mb had good long-term storage stability. The prepared liquid Mb had good immunoreactivity and stability, avoiding storage in freeze-dried powder. It was a promising alternative as the quality control material for Mb detection.</div></div>","PeriodicalId":20421,"journal":{"name":"Practical Laboratory Medicine","volume":"44 ","pages":"Article e00456"},"PeriodicalIF":1.7,"publicationDate":"2025-01-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143140800","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Impact of hemolysis on the levels of proteins associated with aging and age-related neurodegenerative diseases in a multicentric clinical research

IF 1.7 Q3 MEDICAL LABORATORY TECHNOLOGY

Practical Laboratory Medicine

Pub Date : 2025-01-20 DOI: 10.1016/j.plabm.2025.e00455

Masroor Anwar , Km Renu , Abhinay Kumar Singh , Abhilasha Nayal , Bharat Thyagarajan , Peifeng Hu , Jinkook Lee , Sharmistha Dey , A.B. Dey

Introduction

Hemolysis is a known interference factor that has been found to show erroneous effect. Present study analyzes the impact of hemolysis on the concentrations of protein biomarkers of Alzheimer's disease (Aβ42, t-Tau, p-Tau181) along with novel proteins which are currently under investigation (SIRT1,SIRT2,SIRT6,FOXO3A, NFL, Aβ40, GFAP).

Methods

Plasma samples were grouped into two categories: hemolyzed and non-hemolyzed groups. Degree of hemolysis (in percentage) was separately analyzed using Single molecule array (SIMOA) technology. Quantitative analysis for hemolyzed and non-hemolyzed samples were done using surface plasmon resonance (SPR) technology.

Results

The SIMOA analysis indicated that at high levels of hemolysis (1000 mg/dL) there was an increase in NFL protein level up to approximately 30 % whereas p-Tau181 did not show much interference even at higher hemolysate concentration. Aβ40, Aβ42 and GFAP showed modest effect up to hemolysis of 250mg/dL-500 mg/dL. SPR analysis of total Tau (t-Tau), p-Tau181, SIRT1, SIRT6 showed the consistency in the result and there was no significant difference in hemolyzed plasma compared to non-hemolyzed samples. Aβ42 and FOXO3A showed decline in hemolyzed plasma compared to non-hemolyzed samples (4.34 ± 0.18ng/ul; 4.95 ± 0.19ng/ul) and (3.83 ± 0.34ng/ul; 5.12 ± 0.46ng/ul), respectively whereas, a significant increase in the concentration was observed for SIRT2; 2.4 ± 0.10ng/ul in hemolyzed compared to 1.30 ± 0.22ng/ul in non-hemolyzed group.

Conclusions

High grade hemolysis leads to altered protein concentration associated with neurodegeneration. Present study emphasizes the need to have pre-analytical inspection for hemolysis detection especially in a multicentric biomarker study.

{"title":"Impact of hemolysis on the levels of proteins associated with aging and age-related neurodegenerative diseases in a multicentric clinical research","authors":"Masroor Anwar , Km Renu , Abhinay Kumar Singh , Abhilasha Nayal , Bharat Thyagarajan , Peifeng Hu , Jinkook Lee , Sharmistha Dey , A.B. Dey","doi":"10.1016/j.plabm.2025.e00455","DOIUrl":"10.1016/j.plabm.2025.e00455","url":null,"abstract":"<div><h3>Introduction</h3><div>Hemolysis is a known interference factor that has been found to show erroneous effect. Present study analyzes the impact of hemolysis on the concentrations of protein biomarkers of Alzheimer's disease (Aβ42, t-Tau, p-Tau181) along with novel proteins which are currently under investigation (SIRT1,SIRT2,SIRT6,FOXO3A, NFL, Aβ40, GFAP).</div></div><div><h3>Methods</h3><div>Plasma samples were grouped into two categories: hemolyzed and non-hemolyzed groups. Degree of hemolysis (in percentage) was separately analyzed using Single molecule array (SIMOA) technology. Quantitative analysis for hemolyzed and non-hemolyzed samples were done using surface plasmon resonance (SPR) technology.</div></div><div><h3>Results</h3><div>The SIMOA analysis indicated that at high levels of hemolysis (1000 mg/dL) there was an increase in NFL protein level up to approximately 30 % whereas p-Tau181 did not show much interference even at higher hemolysate concentration. Aβ40, Aβ42 and GFAP showed modest effect up to hemolysis of 250mg/dL-500 mg/dL. SPR analysis of total Tau (t-Tau), p-Tau181, SIRT1, SIRT6 showed the consistency in the result and there was no significant difference in hemolyzed plasma compared to non-hemolyzed samples. Aβ42 and FOXO3A showed decline in hemolyzed plasma compared to non-hemolyzed samples (4.34 ± 0.18ng/ul; 4.95 ± 0.19ng/ul) and (3.83 ± 0.34ng/ul; 5.12 ± 0.46ng/ul), respectively whereas, a significant increase in the concentration was observed for SIRT2; 2.4 ± 0.10ng/ul in hemolyzed compared to 1.30 ± 0.22ng/ul in non-hemolyzed group.</div></div><div><h3>Conclusions</h3><div>High grade hemolysis leads to altered protein concentration associated with neurodegeneration. Present study emphasizes the need to have pre-analytical inspection for hemolysis detection especially in a multicentric biomarker study.</div></div>","PeriodicalId":20421,"journal":{"name":"Practical Laboratory Medicine","volume":"44 ","pages":"Article e00455"},"PeriodicalIF":1.7,"publicationDate":"2025-01-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143140814","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

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