Practical Laboratory Medicine最新文献

Development of a rapid LFA test based on direct RT-LAMP for diagnosis of SARS-CoV-2 开发基于直接 RT-LAMP 的快速 LFA 检验，用于诊断 SARS-CoV-2

IF 1.7 Q3 MEDICAL LABORATORY TECHNOLOGY

Practical Laboratory Medicine

Pub Date : 2024-11-01 DOI: 10.1016/j.plabm.2024.e00437

Negar Sadeghi , Neda Shirazi , Moein Dehbashi , Bahareh Maleki , William C. Cho , Zohreh Hojati

Introduction

In response to the rapid spread of the SARS-CoV-2 virus, we developed a rapid molecular approach to diagnose COVID-19 without the need for RNA extraction.

Methods

The study utilized two molecular methods, RT-qPCR and colorimetric RT-LAMP, to diagnose the RdRp and ORF8 genes, respectively, in oro-nasopharyngeal swabs. Due to the high sequence diversity of ORF8 in SARS-CoV and SARS-CoV-2, it has been identified as a suitable target for virus detection. The RT-LAMP method was also carried out directly on heat-treated swab samples. The strip tests were made using gold nanoparticles and combined with the RT-LAMP for further analysis.

Results

The results showed that the isothermal amplification method had a sensitivity of 95 % (95 % C.I.: 86.08 %–98.96 %) and a specificity of 75 % (95 % C.I.: 19.41 %–99.37 %). The RT-LAMP-LFA method was able to distinguish positive and negative samples with 100 % sensitivity (95 % C.I.: 91.96–100) and 77.27 % specificity (95 % C.I.: 54.63–92.18). This method only required heating swab samples for 10 min at 65 °C before the RT-LAMP reaction.

Conclusion

By utilizing the RT-LAMP in combination with the LFA, it is possible to diagnose SARS-CoV-2 rapidly without the need for RNA extraction. The entire process from sample collection to test interpretation takes only 75–90 min, and the results can be interpreted by untrained individuals with the naked eye. By employing the ORF8 gene as a diagnostic target and eliminating the need for RNA extraction, the direct RT-LAMP-LFA method achieves a significant breakthrough that was not previously reported.

方法该研究利用 RT-qPCR 和比色 RT-LAMP 两种分子方法分别诊断口鼻咽拭子中的 RdRp 和 ORF8 基因。由于 ORF8 在 SARS-CoV 和 SARS-CoV-2 中具有高度序列多样性，因此被确定为检测病毒的合适目标。RT-LAMP 方法也是直接在经过热处理的咽拭子样本上进行的。结果表明，等温扩增法的灵敏度为 95 %（95 % C.I.：86.08 %-98.96 %），特异性为 75 %（95 % C.I.：19.41 %-99.37 %）。RT-LAMP-LFA 方法能够区分阳性和阴性样本，灵敏度为 100 %（95 % C.I.：91.96-100），特异度为 77.27 %（95 % C.I.：54.63-92.18）。该方法只需在 RT-LAMP 反应前将拭子样本在 65 °C 下加热 10 分钟即可。从样本采集到检验结果判读的整个过程只需 75-90 分钟，未经训练的人也能用肉眼判读结果。直接 RT-LAMP-LFA 方法采用 ORF8 基因作为诊断靶标，无需提取 RNA，实现了前所未有的重大突破。

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引用次数: 0

The measurement of immunosuppressive drugs by mass spectrometry and immunoassay in a South African transplant setting 在南非移植环境中通过质谱法和免疫测定法测量免疫抑制药物

IF 1.7 Q3 MEDICAL LABORATORY TECHNOLOGY

Practical Laboratory Medicine

Pub Date : 2024-11-01 DOI: 10.1016/j.plabm.2024.e00440

Amy Strydom, Doreen Jacob, Taryn Pillay, Refeletse Malahlela, Sean Currin

Objectives

Liquid chromatography tandem mass spectrometry (LC-MS/MS) is the gold standard for measurement of immunosuppressive drugs (ISDs), but is technically demanding and less accessible in resource-limited countries. Immunoassays can also measure ISD concentrations, but may be limited by cross-reactivity. We evaluated the performance of the Roche electrochemiluminescence immunoassay (ECLIA) for cyclosporine, everolimus and sirolimus against LC-MS/MS in an African population for the first time.

Methods

Bias for ECLIA was estimated by comparing ECLIA-measured ISD concentrations to those obtained by LC-MS/MS in 42, 43 and 47 patient samples for cyclosporine, everolimus and sirolimus, respectively. Precision was assessed by performing replicate measurements of quality control materials.

Results

Deming regression analysis for all ISDs showed strong correlation between ECLIA and LC-MS/MS with a Pearson's r of >0.94. The slopes for cyclosporine, everolimus and sirolimus were 0.94 [95 % CI: 0.87–1.03], 1.35 [95 % CI: 1.23–1.44] and 0.96 [95 % CI: 0.85–1.15] with y-intercepts of 31.60 μg/L [95 % CI: 2.02–57.63], 0.23 μg/L [95 % CI: 0.21 – 0.72] and 2.61 μg/L [95 % CI: 1.30–3.56], respectively. Difference plots showed a median bias of 2.07 % [95 % CI: 1.42 – 6.99 %], 41.2 % [95 % CI: 34.9–51.8 %] and 34.9 % [95 % CI: 28.4–47.3 %] for cyclosporine, everolimus and sirolimus, respectively.

Conclusions

The cyclosporine ECLIA yielded results comparable to LC-MS/MS while poorly comparable results were obtained for everolimus and sirolimus, which may be explained by ISD metabolite cross-reactivity, amongst other factors. The poor comparability, although not unique, is noteworthy and the clinical consequences of these differences require further investigation.

目标液相色谱串联质谱法（LC-MS/MS）是测量免疫抑制剂（ISD）的黄金标准，但技术要求高，在资源有限的国家较难获得。免疫测定也可以测量 ISD 的浓度，但可能会受到交叉反应的限制。我们首次在非洲人群中评估了罗氏电化学发光免疫分析法（ECLIA）与液相色谱-质谱联用法（LC-MS/MS）在环孢素、依维莫司和西罗莫司方面的性能。结果对所有 ISD 进行的回归分析表明，ECLIA 与 LC-MS/MS 之间具有很强的相关性，Pearson's r 为 0.94。环孢素、依维莫司和西罗莫司的斜率分别为 0.94 [95 % CI: 0.87-1.03]、1.35 [95 % CI: 1.23-1.44]和 0.96 [95 % CI: 0.85-1.15]，Y-截距分别为 0.15、0.15 和 0.15。15]，Y-截距分别为 31.60 μg/L [95 % CI：2.02-57.63]、0.23 μg/L [95 % CI：0.21-0.72] 和 2.61 μg/L [95 % CI：1.30-3.56]。差异图显示，环孢素、依维莫司和西罗莫司的中位偏差分别为 2.07 % [95 % CI: 1.42 - 6.99 %]、41.2 % [95 % CI: 34.9 - 51.8 %]和 34.9 % [95 % CI: 28.4 - 47.3 %]。结论环孢素 ECLIA 的检测结果可与 LC-MS/MS 相媲美，而依维莫司和西罗莫司的检测结果可比性较差，这可能与 ISD 代谢物交叉反应等因素有关。可比性差虽然不是独一无二的，但值得注意，这些差异的临床后果需要进一步研究。

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引用次数: 0

Falsely abnormal serum protein electrophoresis after administration of intravenous immunoglobulins (IVIG): A retrospective cohort study 静脉注射免疫球蛋白（IVIG）后血清蛋白电泳异常：回顾性队列研究

IF 1.7 Q3 MEDICAL LABORATORY TECHNOLOGY

Practical Laboratory Medicine

Pub Date : 2024-11-01 DOI: 10.1016/j.plabm.2024.e00434

Andrew Sulaiman, Patrizio Caturegli

Intravenous immunoglobulin (IVIG) therapy, used in several neurologic, hematologic, immunologic and dermatologic conditions, is known to interfere with the results of some serum laboratory tests. We analyzed the potential interference of IVIG on serum protein electrophoresis (SPEP) by reviewing more than a decade of SPEP studies performed by the clinical immunology laboratory of the Johns Hopkins Hospital. Of the total 100,350 SPEP performed between January 1, 2013 and December 31, 2023, 395 contained the keyword IVIG in the pathologist report, contributed by 348 patients confirmed to have received IVIG by chart review. Of the 348 patients, 20 (6 %) had a M-spike on SPEP suggestive of monoclonal gammopathy, while 328 (94 %) did not have it. Of the 20 patients, 14 received IVIG within 30 days from the SPEP collection date, while 6 received beyond 30 days. Serum immunofixation electrophoresis (SIFE) and clinical follow up showed no evidence of monoclonal gammopathy in 5 of the 14 patients. Overall, this 11-year retrospective cohort study showed that 5 of 348 (1.4 %) patients treated with IVIG and tested by SPEP had a false M-spike, that is a spike not confirmed to be caused by a monoclonal gammopathy by subsequent studies. Although small, the false positive rate of 1.4 % suggests that integrating knowledge of recent IVIG administration into the pathologist report would reduce SPEP misdiagnosis.

众所周知，静脉注射免疫球蛋白（IVIG）疗法会干扰某些血清实验室检测的结果，该疗法可用于多种神经、血液、免疫和皮肤病的治疗。我们通过回顾约翰霍普金斯医院临床免疫学实验室十多年来进行的 SPEP 研究，分析了 IVIG 对血清蛋白电泳（SPEP）的潜在干扰。在 2013 年 1 月 1 日至 2023 年 12 月 31 日期间进行的总共 100,350 例 SPEP 中，有 395 例病理学家报告中包含 IVIG 关键字，其中 348 例患者通过病历审查确认接受了 IVIG 治疗。在这 348 名患者中，有 20 人（6%）在 SPEP 中出现了提示单克隆性腺病的 M 峰，而 328 人（94%）没有出现 M 峰。在这 20 名患者中，14 人在 SPEP 采集日期后 30 天内接受了 IVIG 治疗，6 人在 30 天后接受了 IVIG 治疗。血清免疫固定电泳（SIFE）和临床随访显示，14 名患者中有 5 人未发现单克隆抗体病。总体而言，这项为期 11 年的回顾性队列研究显示，在 348 例接受 IVIG 治疗并接受 SPEP 检测的患者中，有 5 例（1.4%）出现了假 M-棘波，即后续研究未证实是由单克隆丙种球蛋白病引起的棘波。1.4% 的假阳性率虽然很小，但表明将近期 IVIG 用药知识纳入病理学家报告可减少 SPEP 误诊。

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引用次数: 0

Glycated albumin in pregnancy correlates negatively with body mass index and contributes to the risk of gestational diabetes mellitus 孕期糖化白蛋白与体重指数呈负相关，并增加妊娠糖尿病的风险

IF 1.7 Q3 MEDICAL LABORATORY TECHNOLOGY

Practical Laboratory Medicine

Pub Date : 2024-10-23 DOI: 10.1016/j.plabm.2024.e00439

Toril Ø. Osestad , Kristin Lilleholt , Øyvind Skadberg , Linda R. Sagedal , Ingvild Vistad , Thomas Hundhausen

Objectives

The aims of our study were to establish a reference interval for glycated albumin (GA) in gestational week 30, to investigate whether GA can replace or reduce the need for oral glucose tolerance test (OGTT) in pregnancy, and to reassess the usefulness of body mass-index (BMI), age and fasting glucose in detection of gestational diabetes (GDM).

Design

and methods: We measured GA in 486 healthy pregnant women. Reference interval was calculated using the central 95 % of the results. ROC curves were created to assess the ability of GA, fasting glucose and BMI separately to detect GDM, and logistic regression analysis was used to estimate risk of developing GDM given the level of the same markers. Finally, multiple logistic regression analysis based on GA, fasting glucose and BMI was used to find a strategy of predicting a patient's risk of GDM.

Results

The reference interval for GA at week 30 of gestation is 6.8–10.3 %. The analysis has a low AUC (0.53) with respect to detecting GDM. It increases slightly to 0.64 when corrected for BMI, as GA is inversely correlated to BMI. Combining GA with fasting glucose and BMI at gestational weeks 16–20 could raise the AUC to 0.80.

Conclusion

GA cannot be recommended to replace OGTT for the diagnosis of GDM. Nor can it be used to identify women at risk of developing GDM. GA combined with fasting glucose and BMI in early pregnancy could be a useful model to estimate risk of GDM.

目的我们的研究旨在确定妊娠 30 周糖化白蛋白（GA）的参考区间，研究 GA 是否可以取代或减少妊娠期口服葡萄糖耐量试验（OGTT），并重新评估体重指数（BMI）、年龄和空腹血糖在检测妊娠糖尿病（GDM）中的作用：我们测量了 486 名健康孕妇的妊娠期糖尿病。以结果的 95% 为中心计算参考区间。绘制了 ROC 曲线，分别评估 GA、空腹血糖和体重指数检测 GDM 的能力，并使用逻辑回归分析估算在相同指标水平下罹患 GDM 的风险。最后，基于 GA、空腹血糖和 BMI 的多重逻辑回归分析被用来寻找预测患者 GDM 风险的策略。该分析在检测 GDM 方面的 AUC 较低（0.53）。由于 GA 与体重指数成反比，因此在对体重指数进行校正后，AUC 略微上升至 0.64。将 GA 与空腹血糖和孕 16-20 周时的 BMI 相结合，可将 AUC 提高到 0.80。GA不能取代OGTT用于诊断GDM，也不能用于识别有患GDM风险的妇女。GA与孕早期空腹血糖和体重指数相结合，可作为估计GDM风险的有用模型。

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引用次数: 0

A novel case of Hb Bart's hydrops fetalis following prenatal diagnosis: Case report from Huizhou, China 产前诊断后出现 Hb Bart 胎儿水肿的新病例：来自中国惠州的病例报告

IF 1.7 Q3 MEDICAL LABORATORY TECHNOLOGY

Practical Laboratory Medicine

Pub Date : 2024-10-23 DOI: 10.1016/j.plabm.2024.e00438

Zeyan Zhong , Dina Chen , Zhiyang Guan , Guoxing Zhong , Zhiyong Wu , Jianmin Chen , Jianhong Chen

Objective

Presentation of a novel case of a patient with Hb Bart's hydrops fetalis, which was accurately identified by SMRT sequencing leading to expand the mutation spectrum of α-thalassemia.

Case report

A 26-year-old pregnant woman and her husband underwent molecular analysis of thalassemia due to abnormal hematological results. The molecular analysis showed that the pregnant woman carried -α^3.7/--^SEA, while her husband exhibited a negative result. Accordingly, the pregnant woman continued the pregnancy until the 19-week gestational age. She was subsequently referred to our department for genetic counseling due to abnormal ultrasound findings in the fetus. A novel deletional α-thal mutation was detected for the husband by MLPA, and the precise location of the mutation was determined through SMRT sequencing, which revealed a 45.2 kb deletion. Later, an interventional umbilical cord blood puncture was offered for the pregnant woman. The cord blood was subjected to capillary electrophoresis, which revealed apparent Hb Bart's and Hb Portland peaks associated with Hb Bart's hydrops fetalis syndrome.

Conclusion

It is imperative that Hb Bart's hydrops fetalis syndrome be diagnosed with the utmost expediency. If results of molecular analysis are not consistent with the clinical hematological findings, the presence of a novel thalassemia could be suspected. To identify the novel genotype, the SMRT sequencing represents an effective method for achieving an accurate diagnosis.

病例报告一位 26 岁的孕妇及其丈夫因血液学结果异常而接受了地中海贫血分子分析。分子分析结果显示，孕妇携带-α3.7/--SEA，而其丈夫的结果为阴性。因此，孕妇继续妊娠至孕 19 周。由于胎儿的超声检查结果异常，她随后被转诊到我科接受遗传咨询。通过 MLPA 检测，发现丈夫有一个新的缺失α-thal 基因突变，并通过 SMRT 测序确定了基因突变的确切位置，发现有一个 45.2 kb 的缺失。随后，为孕妇进行了介入性脐带血穿刺。对脐带血进行了毛细管电泳，发现明显的 Hb Bart's 峰和 Hb Portland 峰与 Hb Bart's 胎儿水肿综合征有关。如果分子分析结果与临床血液学结果不一致，则可能怀疑存在新型地中海贫血。要确定新型基因型，SMRT 测序是实现准确诊断的有效方法。

{"title":"A novel case of Hb Bart's hydrops fetalis following prenatal diagnosis: Case report from Huizhou, China","authors":"Zeyan Zhong , Dina Chen , Zhiyang Guan , Guoxing Zhong , Zhiyong Wu , Jianmin Chen , Jianhong Chen","doi":"10.1016/j.plabm.2024.e00438","DOIUrl":"10.1016/j.plabm.2024.e00438","url":null,"abstract":"<div><h3>Objective</h3><div>Presentation of a novel case of a patient with Hb Bart's hydrops fetalis, which was accurately identified by SMRT sequencing leading to expand the mutation spectrum of α-thalassemia.</div></div><div><h3>Case report</h3><div>A 26-year-old pregnant woman and her husband underwent molecular analysis of thalassemia due to abnormal hematological results. The molecular analysis showed that the pregnant woman carried -α<sup>3.7</sup>/--<sup>SEA</sup>, while her husband exhibited a negative result. Accordingly, the pregnant woman continued the pregnancy until the 19-week gestational age. She was subsequently referred to our department for genetic counseling due to abnormal ultrasound findings in the fetus. A novel deletional α-thal mutation was detected for the husband by MLPA, and the precise location of the mutation was determined through SMRT sequencing, which revealed a 45.2 kb deletion. Later, an interventional umbilical cord blood puncture was offered for the pregnant woman. The cord blood was subjected to capillary electrophoresis, which revealed apparent Hb Bart's and Hb Portland peaks associated with Hb Bart's hydrops fetalis syndrome.</div></div><div><h3>Conclusion</h3><div>It is imperative that Hb Bart's hydrops fetalis syndrome be diagnosed with the utmost expediency. If results of molecular analysis are not consistent with the clinical hematological findings, the presence of a novel thalassemia could be suspected. To identify the novel genotype, the SMRT sequencing represents an effective method for achieving an accurate diagnosis.</div></div>","PeriodicalId":20421,"journal":{"name":"Practical Laboratory Medicine","volume":"42 ","pages":"Article e00438"},"PeriodicalIF":1.7,"publicationDate":"2024-10-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142533310","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

Comparison of qPCR and chromogenic culture methods for rapid detection of group B streptococcus colonization in Vietnamese pregnant women 比较 qPCR 和色原培养法快速检测越南孕妇 B 群链球菌定植情况

IF 1.7 Q3 MEDICAL LABORATORY TECHNOLOGY

Practical Laboratory Medicine

Pub Date : 2024-10-15 DOI: 10.1016/j.plabm.2024.e00435

Manh-Tuan Ha , Huyen Tran-Thi-Bich , Thao Bui-Thi-Kim , My-Linh Nguyen-Thi , Thanh Vu-Tri , Thuy-Duong Ho-Huynh , Tuan-Anh Nguyen

Introduction

Neonatal infections can rapidly become severe, with delays in treatment often proving fatal. Streptococcus agalactiae (Group B Streptococcus, GBS) is a common cause, typically transmitted from colonized pregnant women to neonates during childbirth. In Vietnam, routine prenatal care lacks standardized GBS screening protocols. This study aims to compare enhanced qPCR methods with the culture method, evaluate the diagnostic accuracy of these qPCR procedures, and assess the frequency of GBS infection in pregnant Vietnamese women during their final trimester.

Materials and methods

Pregnant women aged 35 weeks gestation or more were recruited. Rectovaginal swabs were collected and analyzed for GBS using chromogenic culture, a commercial real-time PCR kit, and in-house real-time PCR assays targeting the cfb and sip genes. Clinical diagnostic values were calculated, and GBS prevalence was determined.

Results

The study included 259 pregnant women with a mean age of 30.2 ± 5.0 years. Of these, 96.6 % had gestational ages of 37 weeks or more at delivery. The sensitivity, specificity, positive predictive value, negative predictive value, and accuracy of the cfb-based and sip-based qPCR assays were 94.1/92.7, 99.0/99.5, 97.1/98.5, 97.8/97.3, and 97.6 %, respectively. The Kappa values were excellent (0.940 and 0.939), with results available in under 2 h. GBS prevalence was 24.7 % and 25.5 % by cfb-based and sip-based qPCR assays, aligning with the culture method (25.5 %).

Conclusions

Both direct real-time PCR assays demonstrated high accuracy and were comparable to chromogenic culture in diagnosing GBS. A significant prevalence of GBS colonization was found among Vietnamese pregnant women in their final trimester.

导言：新生儿感染会迅速恶化，延误治疗往往会导致死亡。无乳链球菌（B 组链球菌，GBS）是一种常见的致病菌，通常在分娩过程中由带有菌落的孕妇传染给新生儿。在越南，常规产前护理缺乏标准化的 GBS 筛查方案。本研究旨在比较增强型 qPCR 方法和培养方法，评估这些 qPCR 程序的诊断准确性，并评估越南孕妇在最后三个月感染 GBS 的频率。收集直肠阴道拭子，并使用色原培养法、商用实时 PCR 试剂盒和针对 cfb 和 sip 基因的内部实时 PCR 检测法进行 GBS 分析。计算了临床诊断值，并确定了 GBS 患病率。其中，96.6%的孕妇分娩时胎龄为 37 周或以上。基于 cfb 和基于 sip 的 qPCR 检测的灵敏度、特异性、阳性预测值、阴性预测值和准确性分别为 94.1/92.7、99.0/99.5、97.1/98.5、97.8/97.3 和 97.6%。基于 cfb 和基于 sip 的 qPCR 检测方法的 GBS 感染率分别为 24.7% 和 25.5%，与培养方法（25.5%）一致。结论两种直接实时 PCR 检测方法在诊断 GBS 方面都具有很高的准确性，与色原培养法不相上下。

{"title":"Comparison of qPCR and chromogenic culture methods for rapid detection of group B streptococcus colonization in Vietnamese pregnant women","authors":"Manh-Tuan Ha , Huyen Tran-Thi-Bich , Thao Bui-Thi-Kim , My-Linh Nguyen-Thi , Thanh Vu-Tri , Thuy-Duong Ho-Huynh , Tuan-Anh Nguyen","doi":"10.1016/j.plabm.2024.e00435","DOIUrl":"10.1016/j.plabm.2024.e00435","url":null,"abstract":"<div><h3>Introduction</h3><div>Neonatal infections can rapidly become severe, with delays in treatment often proving fatal. <em>Streptococcus agalactiae</em> (Group B <em>Streptococcus</em>, GBS) is a common cause, typically transmitted from colonized pregnant women to neonates during childbirth. In Vietnam, routine prenatal care lacks standardized GBS screening protocols. This study aims to compare enhanced qPCR methods with the culture method, evaluate the diagnostic accuracy of these qPCR procedures, and assess the frequency of GBS infection in pregnant Vietnamese women during their final trimester.</div></div><div><h3>Materials and methods</h3><div>Pregnant women aged 35 weeks gestation or more were recruited. Rectovaginal swabs were collected and analyzed for GBS using chromogenic culture, a commercial real-time PCR kit, and in-house real-time PCR assays targeting the <em>cfb</em> and <em>sip</em> genes. Clinical diagnostic values were calculated, and GBS prevalence was determined.</div></div><div><h3>Results</h3><div>The study included 259 pregnant women with a mean age of 30.2 ± 5.0 years. Of these, 96.6 % had gestational ages of 37 weeks or more at delivery. The sensitivity, specificity, positive predictive value, negative predictive value, and accuracy of the <em>cfb</em>-based and sip-based qPCR assays were 94.1/92.7, 99.0/99.5, 97.1/98.5, 97.8/97.3, and 97.6 %, respectively. The Kappa values were excellent (0.940 and 0.939), with results available in under 2 h. GBS prevalence was 24.7 % and 25.5 % by <em>cfb</em>-based and <em>sip</em>-based qPCR assays, aligning with the culture method (25.5 %).</div></div><div><h3>Conclusions</h3><div>Both direct real-time PCR assays demonstrated high accuracy and were comparable to chromogenic culture in diagnosing GBS. A significant prevalence of GBS colonization was found among Vietnamese pregnant women in their final trimester.</div></div>","PeriodicalId":20421,"journal":{"name":"Practical Laboratory Medicine","volume":"42 ","pages":"Article e00435"},"PeriodicalIF":1.7,"publicationDate":"2024-10-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142445068","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

The interference and elimination of nitrite on determination of total urinary protein by Pyrogallol Red–Molybdate method 焦红-钼酸盐法测定尿总蛋白时亚硝酸盐的干扰与消除

IF 1.7 Q3 MEDICAL LABORATORY TECHNOLOGY

Practical Laboratory Medicine

Pub Date : 2024-10-15 DOI: 10.1016/j.plabm.2024.e00436

Shijian Fan , Jiaqing Liu , Yu Wei , Jie Yao , Juan Cheng , Yang Tong , Qiang Zhou , Yuanhong Xu

Background

Some urine samples showed negative values of total urinary protein by Pyrogallol red–molybdate (PRM) method. Interestingly, these samples showed notably high levels of nitrite in the urine dipstick test.

Methods

A total of 120 urine samples were collected and categorized into four groups (0 - < 100, 100-<500, 500-<1000, ≥1000 mg/L) based on total urinary protein concentration. Various concentrations of nitrite (2, 10, 50, 100, and 200 mg/L) were added to urine samples to investigate potential interference of nitrite in total urinary protein measurement by the PRM method. Additionally, different concentrations of L-ascorbic acid (100, 500, 1000, and 2000 mg/L) were added to urine-nitrite mixtures to explore the possibility of reversing the interference effects.

Results

2 mg/L nitrite had no impact on the test results.10 mg/L nitrite only showed significant effects on the detection results among the groups with 0 - < 100 and 100-<500 mg/L (P < 0.05). 200 mg/L nitrite caused a noticeable decrease in the urinary protein detection results of the four groups (0 - < 100, 100-<500, 500-<1000, ≥1000 mg/L), and the concentrations were reduced to 2.7 %, 26.85 %, 75.22 %, and 89.33 % of their original levels, respectively. 500 mg/L L-ascorbic acid effectively eliminated the interference from 200 mg/L nitrite and almost had no effect on detection.

Conclusions

Nitrite had negative effect on the detection of total urinary protein by PRM method and L-ascorbic acid was effective in counteracting the interference caused by nitrite.

背景部分尿样经焦聚醇红钼酸盐（PRM）法检测显示尿总蛋白呈阴性。方法共收集了 120 份尿液样本，并根据尿液总蛋白浓度分为四组（0 - 100、100-500、500-1000、≥1000 mg/L）。在尿样中加入不同浓度的亚硝酸盐（2、10、50、100 和 200 mg/L），以研究亚硝酸盐对 PRM 方法测定尿蛋白总量的潜在干扰。结果2 毫克/升亚硝酸盐对检测结果没有影响。10 毫克/升亚硝酸盐只对 0 - < 100 和 100 - < 500 毫克/升组的检测结果有显著影响（P < 0.05）。200 mg/L 亚硝酸盐使四组（0 - <100、100-<500、500-<1000、≥1000 mg/L）的尿蛋白检测结果明显下降，浓度分别降至原来的 2.7 %、26.85 %、75.22 % 和 89.33 %。结论亚硝酸盐对 PRM 法检测尿总蛋白有负面影响，L-抗坏血酸能有效抵消亚硝酸盐的干扰。

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引用次数: 0

Validation of a LC-MS/MS assay for citric acid, cysteine and oxalic acid determination and its application to explore pre-analytical sample storage 验证测定柠檬酸、半胱氨酸和草酸的 LC-MS/MS 分析法及其在探索分析前样品储存中的应用

IF 1.7 Q3 MEDICAL LABORATORY TECHNOLOGY

Practical Laboratory Medicine

Pub Date : 2024-09-30 DOI: 10.1016/j.plabm.2024.e00433

Ying Shen , Xia Luo , Qing Guan , Wenjie Lou , Liming Cheng

Objectives

Citrate, oxalate and cystine in 24-h urine are considered to be associated with the incidence and recurrence risk of urinary stone disease (USD). An evaluation of the LC-MS/MS kit for simultaneous quantification of the three analytes was undertaken.

Design

& Methods: The analytical performance of the kit was investigated based on FDA, EMA and CLSI guidelines. To promote the standardization of sample storage, this kit has been applied to perform systematic pre-analytical stability study of these analytes in urine.

Results

This method was validated with good linearity with accuracy of 93.1%–104 %. Intra-day and inter-day imprecision were ≤5.55 % and 5.34 %, respectively. Recoveries of citrate, oxalate and cystine added to clinical samples were in the range of 92.0–103 %, 94.8–100 % and 99.0–107 % with CV ≤ 5.52 %. It was recommended that urine preserved with hydrochloric acid could be preferable in consideration of both reliable test results and neglected sample heterogeneity.

Conclusions

This kit is suitable for measurement of citrate, oxalate and cystine for understanding the etiology of urinary stones, and the proper storage of urine samples is crucial for the correctness of the test results.

目的 24 小时尿液中的柠檬酸盐、草酸盐和胱氨酸被认为与尿石症（USD）的发病率和复发风险有关。我们对 LC-MS/MS 试剂盒进行了评估，以同时定量分析这三种分析物：根据 FDA、EMA 和 CLSI 指南对试剂盒的分析性能进行了调查。结果：该方法线性关系良好，准确度为 93.1%-104%。日内和日间误差分别小于5.55%和5.34%。临床样本中柠檬酸盐、草酸盐和胱氨酸的回收率分别为 92.0%-103%、94.8%-100% 和 99.0%-107%，CV ≤ 5.52%。结论：该试剂盒适用于测定枸橼酸盐、草酸盐和胱氨酸，可用于了解尿路结石的病因，尿样的妥善保存对检测结果的正确性至关重要。

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引用次数: 0

Performance evaluation of Eu3+-based CRP/SAA and PCT/IL-6 lateral flow immunoassay kits and their diagnostic value in respiratory tract infections 基于 Eu3+ 的 CRP/SAA 和 PCT/IL-6 侧流免疫测定试剂盒的性能评估及其在呼吸道感染中的诊断价值

IF 1.7 Q3 MEDICAL LABORATORY TECHNOLOGY

Practical Laboratory Medicine

Pub Date : 2024-09-27 DOI: 10.1016/j.plabm.2024.e00432

Mingxin Lin , Jing Zhang , Jianxing Cai , Jumei Liu , Min Zhu , Ke Li , Miaoyun Hu , Chenxi Li , Huiming Ye

Objectives

Respiratory infections are among the most common infectious diseases, resulting in significant morbidity and mortality. C-reactive protein (CRP), serum amyloid A (SAA), procalcitonin (PCT), and interleukin-6 (IL-6) are advantageous for diagnosing respiratory tract infections. This study assessed the analytical performance and accuracy of new kits for Eu3⁺-based CRP/SAA and PCT/IL-6 lateral flow immunoassay and its diagnostic value in respiratory tract infections.

Methods

This study evaluated the detection performance of a test kit using guidelines from the Center for Medical Device Evaluation (CMDE) and the Clinical and Laboratory Standards Institute (CLSI). The test results were compared to those of the commercial kits (CRP: Mindray; SAA: Norman; PCT: Shanghai Upper; IL-6: Wantai BioPharm). A total of 156 patients with respiratory tract infections (53 with bacterial infections (Bac group); 50 with viral infections (Vir group); and 53 with co-infections (Bac + Vir group)) were enrolled, along with 50 healthy controls (HC group). Venous blood samples were collected to measure levels of SAA, PCT, CRP, and IL-6 using both the test and commercial kits. The diagnostic value of these biomarkers was assessed using receiver operating characteristic (ROC) curves.

Results

Correlation analysis demonstrated a strong concordance between the test kits and commercial kits (CRP: r = 0.9396, P < 0.0001; SAA: r = 0.8986, P < 0.0001; PCT: r = 0.9594, P < 0.0001; IL-6: r = 0.9009, P < 0.0001). The diagnostic performance of the test kits in identifying bacterial, viral, and co-infections was highly consistent with that of the commercial kit.

Conclusions

The Eu3⁺-based CRP/SAA and PCT/IL-6 lateral flow immunoassay test kits demonstrated high levels of consistency with commercial kits in terms of quantitative outcomes and diagnostic performance for respiratory tract infections.

目的呼吸道感染是最常见的传染病之一，会导致严重的发病率和死亡率。C反应蛋白（CRP）、血清淀粉样蛋白A（SAA）、降钙素原（PCT）和白细胞介素-6（IL-6）是诊断呼吸道感染的有利指标。本研究评估了基于 Eu3+ 的 CRP/SAA 和 PCT/IL-6 侧流免疫测定新试剂盒的分析性能和准确性及其在呼吸道感染中的诊断价值。检测结果与商用试剂盒（CRP：Mindray；SAA：Norman；PCT：Shanghai Upper；IL-6：Wantai BioPharm）的检测结果进行了比较。共招募了 156 名呼吸道感染患者（53 名细菌感染患者（Bac 组）；50 名病毒感染患者（Vir 组）；53 名合并感染患者（Bac + Vir 组））和 50 名健康对照组（HC 组）。采集静脉血样本，使用检测试剂盒和商业试剂盒测量 SAA、PCT、CRP 和 IL-6 的水平。结果相关性分析表明，检测试剂盒与商用试剂盒之间具有很强的一致性（CRP：r = 0.9396，P <；0.0001；SAA：r = 0.8986，P <；0.0001；PCT：r = 0.9594，P <；0.0001；IL-6：r = 0.9009，P <；0.0001）。结论基于 Eu3+ 的 CRP/SAA 和 PCT/IL-6 侧流免疫测定试剂盒在呼吸道感染的定量结果和诊断性能方面与商业试剂盒具有高度的一致性。

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引用次数: 0

The potential relationship between EasyNAT system Tt values and Cobas z480 Ct values in the detection of SARS-Cov-2 EasyNAT 系统 Tt 值与 Cobas z480 Ct 值在检测 SARS-Cov-2 中的潜在关系

IF 1.7 Q3 MEDICAL LABORATORY TECHNOLOGY

Practical Laboratory Medicine

Pub Date : 2024-09-26 DOI: 10.1016/j.plabm.2024.e00431

Lu-Qing Zheng, Qing-Yong Wang

Objectives

This study aimed to evaluate the potential relationship between the time threshold (Tt) values of a commercial EasyNAT system, which is based on cross priming amplification (CPA) technology, and the cycle threshold (Ct) values of the Cobas z480 analyzer, which is based on a real-time fluorescence polymerase chain reaction (PCR) method, in the detection of severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) from oropharyngeal swabs. Design and Methods: Data were retrospectively collected from a clinical laboratory between December 4, 2022 and July 1, 2024.

Results

A total of 277 EasyNAT-positive samples (Tt values from 3.83 to 29.5) were simultaneously investigated using the Cobas z480 analyzer (Ct values from 10.74 to 38.78). The concordance rate between the two systems was 100 %. Among the positive samples, the mean and maximum PCR Ct values of O and N genes increased in line with increasing Tt values of the left and right amplification areas of the EasyNAT system. The maximum Ct values of the O or N gene determined by the Cobas z480 analyzer were no more than 29.52 when the Tt values of the left or right amplification areas of the UC0116 analyzer were no more than 6.

Conclusions

The safe, simple, fast, accurate, and automatic EasyNAT system used in conjunction with a PCR system might be a better choice for the detection of SARS-CoV-2 in hospitals, especially in settings without sophisticated PCR facilities. The Tt value (≤6) of the EasyNAT system can be a reference index for estimating the maximum Ct value (29.52) in SARS-CoV-2-positive samples.

目的本研究旨在评估基于交叉引物扩增（CPA）技术的商用 EasyNAT 系统的时间阈值（Tt）与基于实时荧光聚合酶链反应（PCR）方法的 Cobas z480 分析仪的周期阈值（Ct）之间在检测口咽拭子中的严重急性呼吸系统综合征冠状病毒-2（SARS-CoV-2）方面的潜在关系。设计与方法：结果使用 Cobas z480 分析仪同时检测了 277 份 EasyNAT 阳性样本（Tt 值从 3.83 到 29.5 不等）（Ct 值从 10.74 到 38.78 不等）。两个系统的吻合率为 100%。在阳性样本中，O 和 N 基因的平均 PCR Ct 值和最大 PCR Ct 值随着 EasyNAT 系统左右扩增区 Tt 值的增加而增加。当 UC0116 分析仪左右扩增区的 Tt 值不超过 6 时，Cobas z480 分析仪测定的 O 或 N 基因的最大 Ct 值不超过 29.52。EasyNAT 系统的 Tt 值（≤6）可以作为估计 SARS-CoV-2 阳性样本中最大 Ct 值（29.52）的参考指标。

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引用次数: 0