Interferon-gamma release assays as a tool for differential diagnosis of gastrointestinal tuberculosis

In this editorial,we comment on an article published in a recent issue of the World Journal of Clinical Cases.There is a pressing need for reliable tools for diagnosing tuberculosis(TB)of the gastrointestinal tract.Despite advancements in the diagnosis and treatment,TB remains a global health challenge.Ali et al demon-strated that TB may mimic gastrointestinal conditions,such as gastric outlet obstruction,causing a delay in the diagnosis.Furthermore,the latter complication is frequently observed during infections,including Helicobacter pylori,and rarely is related to TB,as in the presented case.In line with this,we think that laboratory tests based on interferon-gamma release assays can be a helpful tool for diagnosing latent TB paced in the gastrointestinal tract.Innovative strategies and approaches for diagnosing latent/active extra pulmonary TB are crucial for establishing the diagnosis early and enhancing treatment strategies to mitigate the global burden of TB.

Interferon-gamma release assays in tuberculous uveitis:a comprehensive review

·Tuberculous uveitis(TBU)comprises a broad clinical spectrum of ocular manifestations,making its diagnosis challenging.Ophthalmologists usually require evidence from investigations to confirm or support a clinical diagnosis of TBU.Since direct isolation of the causative organism from ocular specimens has limitations owing to the small volume of the ocular specimens,resultant test positivities are low in yield.Immunodiagnostic tests,including the tuberculin skin test and interferon-gamma release assays(IGRAs),can help support a clinical diagnosis of TBU.Unlike the tuberculin skin test,IGRAs are in vitro tests that require a single visit and are not affected by prior Bacillus Calmette-Guerin vaccination.Currently,available IGRAs consist of different techniques and interpretation methods.Moreover,newer generations have been developed to improve the sensitivity and ability to detect active tuberculosis.This narrative review collates salient practice points as a reference for general ophthalmologists,such as evidence for the utilization of IGRAs in patients with suspected TBU,and summarizes basic knowledge and details of clinical applications of these tests in a clinical setting.

Evaluation of Interferon-Gamma Release Assay Testing and Tuberculin Skin Test for Early Diagnosis of Tuberculosis in Children and Adolescents

Background: This study aimed to evaluate the diagnostic value of interferon-γ release assay (IGRA), a sensitive microbiological diagnostic method, in children and adolescents with suspected tuberculosis in a country with a high burden of tuberculosis. Method: This study included 581 children and adolescents aged 4 - 19 years who were suspected of having tuberculosis, were latently infected with Mycobacterium tuberculosis, and had received at least one dose of BCG vaccine between April 17, 2019, and February 24, 2021. The study evaluated the TST results of 106 patients who had a positive Quantiferon test and were suspected of having tuberculosis. Results: The study included 581 patients aged between 4 and 19 years. Of these, 106 patients tested positive for the Quantiferon test, while 19 were indeterminate and 456 were negative. The Quantiferon test positivity rate was 18.24%. Among the 106 QFT-Plus-positive cases, 23 patients also tested positive for TST. The difference in distribution was found to be statistically significant. Conclusion: The QFT-Plus test is considered an alternative to TST and other microbiological diagnostic methods for early tuberculosis diagnosis, particularly in children and adolescents.

Performance and correlation of interferon gamma release assays and tuberculin skin test in HIV-infected children and adolescents with immune reconstitution

Objective:To evaluate the performance of interferon gamma release assays and tuberculin skin test in HIV-infected children and adolescents with immune reconstitution.Methods:A cross-sectional study was conducted in HIV-infected patients aged 5-18 years receiving antiretroviral treatment with CD4 T-lymphocytes>25%or>500 cells/mm3 for at least 6 months.QuantiF ERON-TB Gold,T-SPOT.TB,and tuberculin skin test were performed in each patient.Results:A total of 50 patients were enrolled with median age of 13.7 years,CD4 counts of 753(IQR:587-989)cells/mm3.Among 27 patients with tuberculosis(16)or tuberculosis exposure(11),8(29.6%)were positive to at least one test,2(7.4%)were positive QuantiFERON-TB Gold,3(11.1%)positive T-SPOT.TB,and 7(25.9%)had tuberculin skin test≥5 mm.Among 23 patients without history of tuberculosis or exposure,all had negative interferon gamma release assays,while 2(8.7%)had positive tuberculin skin test.Conclusions:All tests had low sensitivity despite immune reconstitution.

结核特异性IL-2释放试验在结核感染中的诊断价值探讨

目的基于结核特异性白细胞介素2(IL-2)释放试验的结果,探讨IL-2在辅助诊断活动性结核(ATB)感染中的作用,以及用于监测治疗效果的价值。方法回顾性分析临床进行了结核感染排查的病例599例,其中确诊为活动性结核感染的病例128例,除外活动性结核感染的病例471例。(1)对与阴性对照共同孵育后上清中IL-2(N)检测值制作ROC工作曲线,分析失去抗原刺激后PBMCs持续释放IL-2的能力;(2)对抗原刺激后的响应值IL-2(T-N),制作ROC工作曲线,分析对活动性结核感染的诊断价值;(3)对比分析两组患者IL-2的阳性率;(4)观察响应值IL-2(T-N)阴性和阳性转化时间与疾病进程的依从性。结果IL-2(N)ROC曲线下面积为0.460,95%CI 0.402~0.516;IL-2(T-N)ROC曲线下面积为0.788,95%CI 0.745~0.832;在活动性结核感染组,IL-2(T-N)的阳性率显著高于非活动性结核感染组(χ^(2)=110.858,P<0.001);IL-2发生阴阳性转化的平均时间为3.1个月,时间区间为0.5~7个月。结论PBMCs失去抗原刺激后IL-2仅低水平分泌,对诊断ATB没有价值;结核分枝杆菌特异性抗原刺激后的响应值IL-2(T-N),用于诊断ATB具有一定准确性;IL-2(T-N)阴阳性转化的时间较短,在监测治疗效果方面存在一定价值,监测时间间隔建议为3个月。

Tuberculosis in kidney transplant candidates and recipients

Tuberculosis(TB)is the leading cause of infectious mortality and morbidity in the world,second only to coronavirus disease 2019.Patients with chronic kidney disease and kidney transplant recipients are at a higher risk of developing TB than the general population.Active TB is difficult to diagnose in this population due to close mimics.All transplant candidates should be screened for latent TB infection and given TB prophylaxis.Patients who develop active TB pre-or post-trans-plantation should receive multidrug combination therapy of antitubercular therapy for the recommended duration with optimal dose modification as per glomerular filtration rate.

中国肾移植受者结核病临床诊疗指南(2023版)

本指南旨在为肾移植受者结核病的临床管理提供全面而实用的指导。首先,概述了肾移植受者结核病的特殊性,强调了其高发生率及临床表现的多样性。为了更好地理解患者的病情,建议在移植前进行结核病相关的评估,并注意移植术后对结核病的监测。在诊断方面,详细介绍了目前常用的结核病诊断方法,并提供了在肾移植受者中的适用性评估。在确诊后,讨论了在免疫抑制药应用的背景下,如何平衡结核病治疗和排斥反应的策略,并关注了潜在的药物相互作用。预防方面,强调了在肾移植前对结核病的筛查。本指南旨在提高医务人员对肾移植受者结核病管理的认知,促进更有效的临床实践,提高受者的生活质量。

Early screening to identify and diagnose primary nasal tuberculosis in patients with tumor necrosis factor inhibitors

In this editorial,we comment on the article by Liu et al.Based on our analysis of a case report,we consider that early screening and recognition of primary nasal tuberculosis are crucial for patients undergoing treatment with tumor necrosis factor inhibitor(TNFi).While TNFi therapy increases the risk of reactivating latent tuberculosis,primary nasal tuberculosis remains rare due to the protective mechanisms of the nasal mucosa.Risk factors for primary nasal tuberculosis include minimally invasive nasal surgery,diabetes,and human immunodefi ciency virus.Patients with early symptoms such as nasal congestion,rhinorrhea,altered olfaction,epistaxis,or ulceration,and unresponsive to conventional antibiotics and antihistamines should undergo early rhinoscopy,possibly followed by repeated tissue biopsies and acid-fast bacilli culture when necessary.When diagnosis is challenging,it is essential to consider local tuberculosis epidemiology and the efficacy of diagnostic antituberculosis treatment.The preferred method for tuberculosis screening is the Interferon Gamma Release Assay,with a general recommendation for screening at 3 and 6 months after initial treatment and then every six months.However,the optimal frequency is not yet consensus-driven and may be increased in economically viable settings.

结核病γ干扰素释放试验刺激抗原研究进展

结核分支杆菌复合群引起的人和动物结核病,对人类健康及畜牧业发展造成了严重的威胁,早期诊断对于结核病的防控至关重要。在结核病众多诊断方法中,免疫学诊断方法中γ干扰素释放试验具有很好的应用前景。γ干扰素释放试验刺激抗原以结核菌素(purified protein derivative,PPD)为主,然而PPD抗原成分复杂,诊断特异性和敏感性有待提高,因此寻找可以代替PPD的特异性刺激抗原成为了研究热点。论文系统综述了近年来报道用于γ干扰素释放试验的刺激抗原,重点概述了刺激抗原的免疫特性、刺激效果以及各类刺激抗原的优势、局限性和应用前景,以期为人和动物结核病的诊断及防控提供参考。

3种检测方法对结核病诊断效能的比较

目的评价γ-干扰素释放试验(IGRA)、涂片抗酸染色镜检(AFB)和实时荧光定量核酸扩增检测技术(Xpert MTB/RIF)诊断结核病的应用价值。方法选取2019年6月至2023年6月在陕西省结核病防治研究所就诊的疑似结核病患者397例作为研究对象,所有研究对象均同时行IGRA、AFB、Xpert MTB/RIF检测,收集患者的检测结果和病例资料。排除无法诊断是否为结核病的患者39例,最终纳入358例,并根据临床诊断结果将其分为结核患者141例和非结核患者217例。比较IGRA、AFB、Xpert MTB/RIF在结核病中的阳性检出情况及其对结核病的诊断效能。结果IGRA、AFB、Xpert MTB/RIF检测结核病的灵敏度分别为95.74%、31.21%、56.74%,IGRA的灵敏度高于AFB和Xpert MTB/RIF(χ^(2)=126.66,P<0.01;χ^(2)=59.22,P<0.01),Xpert MTB/RIF的灵敏度高于AFB(χ^(2)=18.65,P<0.01);IGRA、AFB、Xpert MTB/RIF检测结核病的特异度分别为60.37%、99.54%、100.00%,其中AFB和Xpert MTB/RIF的特异度均高于IGRA(χ^(2)=103.86,P<0.01;χ^(2)=107.25,P<0.01),而AFB和Xpert MTB/RIF的特异度比较,差异无统计学意义(χ^(2)=1.00,P=0.32)。IGRA、AFB、Xpert MTB/RIF检测结核病的准确率分别为74.30%、72.63%、82.96%,其中Xpert MTB/RIF的准确率最高,且3种检测方法的准确率比较,差异有统计学意义(χ^(2)=12.30,P<0.01)。IGRA与AFP检测结果的总符合率为31.21%(44/141),阳性符合率为95.45%(42/44),阴性符合率为4.12%(4/97);Xpert MTB/RIF与AFP检测结果的总符合率为74.47(105/141),阳性符合率为100.00%(44/44),阴性符合率为62.89%(61/91)。结论在结核病诊断中,Xpert MTB/RIF和AFB的特异度高于IGRA,而IGRA和Xpert MTB/RIF的灵敏度高于AFB,临床可根据3种方法各自的优势应用于结核病的诊断,以此来提高结核病的临床诊治效率。

Xpert、TB-IGRA及CT诊断脊柱结核的价值

目的:探讨Xpert、TB-IGRA及CT对脊柱结核的诊断价值及CT检查病变情况。方法:选取89例疑似脊柱结核患者,均行病理、Xpert、TB-IGRA及CT检查,统计病理结果,比较Xpert、TB-IGRA、CT与病理结果,分析Xpert、TB-IGRA、CT对脊柱结核的诊断价值,并对比CT与病理检查脊柱结核病变情况。结果:89例疑似脊柱结核患者病理结果显示,阳性60例,阴性29例。CT诊断脊柱结核的阴性预测值、准确度高于Xpert;CT诊断脊柱结核的AUC值高于Xpert、TB-IGRA,且TB-IGRA高于Xpert(P<0.05)。CT检查脊柱结核病变为骨质破坏、韧带下播散占比与病理结果比较有差异(P<0.05)。结论:CT诊断脊柱结核的价值较高,对骨质破坏、韧带下播散的检出情况较好。

干扰素体外释放酶联免疫检验在结核病临床诊断中的应用

目的探讨干扰素体外释放酶联免疫检验(IGRA)在结核病临床诊断中的应用价值。方法选择2022年9月—2023年9月在菏泽市传染病医院经初步检查怀疑为结核病的530例患者作为研究对象,对所有患者实施IGRA和结核菌素皮肤试验,以结核杆菌DNA检测结果作为诊断的“金标准”,比较应用IGRA与结核菌素皮肤试验诊断结核病所得诊断结果,分析两种方法诊断结核病的结果与结核杆菌DNA检测结果的一致性。结果在530例疑似结核病患者中,经结核杆菌DNA检测共412例患者确诊结核病(包括肺结核329例,肺外结核83例),检出率为77.74%。IGRA诊断结核病的敏感度、特异度、阳性预测值、阴性预测值、准确度均显著高于结核菌素皮肤试验(敏感度:98.30%比95.15%;特异度:97.46%比88.98%;阳性预测值:99.26%比96.79%;阴性预测值:94.26%比84.00%;准确度:98.11%比93.77%;均P<0.05)。IGRA和结核菌素皮肤试验诊断结核病的结果与结核杆菌DNA检测结果的一致性分析表明,IGRA与结核杆菌DNA检测结果高度一致(κ值为0.894),结核菌素皮肤试验与结核杆菌DNA检测结果中度一致(κ值为0.657)。结论在对结核病进行诊断时,应用IGRA进行检测具有较高的诊断价值,可实现对结核病的灵敏和准确诊断。

In vitro antihistamine-releasing activity of a peptide derived from wasp venom of Vespa orientalis

Objective: To investigate the antihistamine-releasing effect of a peptide isolated from wasp venom of Vespa orientalis.Methods: This peptide was separated from crude venom by chromatography methods and mass spectrometry. Then various concentrations(2, 4, 8, 16, 32, 64, 128 and256 mmol/L) of the peptide were incubated with mast cells and lactate dehydrogenase assay was performed.Results: No significant effect was observed in lactate dehydrogenase absorbance under128 mmol/L concentration. This implied that the peptide did not cause cell death in mast cells and consequently, histamine release did not happen. Moreover, the results showed the IC50 of mast cells degranulation at 126 mmol/L, which was approximately high implying that this peptide had high selectivity for normal cells and did not cause histamine release from these cells.Conclusions: This would be a great aim in new drug development, in which an agent acts potentially on its target tissue without activating the immune system.

UHPLC-MS/MS法测定盐酸普萘洛尔缓释片中基因毒性杂质N-亚硝基普萘洛尔

目的建立盐酸普萘洛尔缓释片中基因毒性杂质N-亚硝基普萘洛尔的超高效液相色谱-串联质谱(UHPLC-MS/MS)检测方法。方法Waters ACQUITY UPLC CSH TM C 18色谱柱(3.0 mm×150 mm,1.7μm),10 mmol·L^(-1)甲酸铵的水溶液(含0.1%甲酸)作为流动相A,乙腈溶液(含0.1%甲酸)作为流动相B,梯度洗脱,流速为0.5 mL·min^(-1),柱温为50℃,进样器温度为5℃,进样体积为10μL,采用多反应监测(MRM)模式,对盐酸普萘洛尔缓释片中的N-亚硝基普萘洛尔进行定量检测。结果N-亚硝基普萘洛尔在1~20 ng·mL^(-1)范围内具有良好的线性关系。低、中、高3个浓度的加样回收率(n=3)在98.4%~103.2%之间,RSD≤2.7%。检测限和定量限分别为0.09 ng·mL^(-1)和0.3 ng·mL^(-1)。检出盐酸普萘洛尔缓释片中基因毒性杂质N-亚硝基普萘洛尔含量为1.8μg·g^(-1)。结论该方法灵敏度高、专属性强,可用于测定盐酸普萘洛尔缓释片中的N-亚硝基普萘洛尔,为盐酸普萘洛尔缓释片的质量控制提供参考。

抗酸染色、TB-IGRA与Xpert MTB/RIF联合检测在结核病中的诊断价值

目的探究抗酸染色、结核分枝杆菌γ-干扰素体外释放试验(TB-IGRA)与结核分枝杆菌及利福平耐药实时荧光定量核酸扩增(Xpert MTB/RIF)联合检测在结核病中的诊断价值。方法回顾性分析2020年1月至2021年12月接收的338例结核病确诊患者和60例健康体检人员的抗酸染色、TB-IGRA和Xpert MTB/RIF实验室检测结果。比较抗酸染色、TB-IGRA、Xpert MTB/RIF单项及联合检测对结核病的诊断结果;以结核分枝杆菌培养结果为诊断金标准,比较抗酸染色、TB-IGRA、Xpert MTB/RIF单项及联合检测对结核病的诊断效能。结果Xpert MTB/RIF对结核病的阳性符合率为79.0%(267/338),明显高于抗酸染色的57.1%(193/338)、TB-IGRA的68.3%(231/338)(P<0.05);抗酸染色对结核病的阳性符合率低于TB-IGRA(P<0.05)。三者联合检测诊断结核病的灵敏度、准确度、阴性预测值明显高于抗酸染色、TB-IGRA、Xpert MTB/RIF、抗酸染色+TB-IGRA、抗酸染色+Xpert MTB/RIF(P<0.05);不同方法诊断结核病的特异度和阳性预测值比较,差异无统计学意义(P>0.05)。结论TB-IGRA和Xpert MTB/RIF在结核病检测方面优势明显大于抗酸染色,三者联合检测可显著提高结核病的诊断效能,值得临床推广。

HIV/AIDS患者分枝杆菌检出情况分析

目的分析人类免疫缺陷病毒(HIV)感染/获得性免疫缺陷综合征(AIDS)患者合并分枝杆菌感染的实验室检出情况。方法收集2018年1月—2021年12月首都医科大学附属北京佑安医院HIV/AIDS患者送检的1379例分枝杆菌抗酸染色样本、1975例分枝杆菌培养样本、1406例结核分枝杆菌(MTB)聚合酶链反应(PCR)检测样本和522例γ-干扰素释放试验(IGRA)样本实验室检测结果,分析不同检测方法分枝杆菌阳性检出情况和联合检出情况。结果抗酸染色、培养、MTB-PCR、IGRA分枝杆菌阳性率分别为3.41%、5.47%、5.83%、11.49%。同时送检抗酸染色和MTB-PCR或IGRA时,抗酸染色阳性率有所上升;同时送检培养和MTB-PCR或IGRA时,培养法阳性率有所上升;4种方法均送检时阳性率高,且易鉴别出MTB和非结核分枝杆菌(NTM)。MTB-PCR和IGRA为鉴定MTB的特异性方法,2种方法鉴定结果一致性较好。结论抗酸染色法和培养法检测HIV/AIDS患者合并分枝杆菌感染阳性率较低,PCR和IGRA可有效鉴别MTB和NTM。临床高度怀疑HIV/AID患者分枝杆菌感染时,建议多种方法联合检测。

化学发光法干扰素-γ释放试验在肺结核辅助诊断中的价值

目的探讨化学发光法干扰素-γ释放试验在肺结核辅助诊断中的临床应用价值。方法选取2019年3月1日—2020年12月31日潍坊市人民医院183例肺结核患者和33名健康体检者(对照组);将183例肺结核患者分为活动性肺结核组(109例)和非活动性肺结核组(74例)。对所有研究对象行化学发光干扰素-γ释放试验、结核菌素皮内试验、T-SPOT TB试验和血常规检测,比较不同方法诊断肺结核的效能和活动性肺结核组干扰素-γ释放试验阳性、阴性患者血常规检测结果的差异。结果活动性肺结核组、非活动性肺结核组、对照组化学发光干扰素-γ释放试验阳性率分别为92.66%、62.16%、9.09%,3个组阳性率差异有统计学意义(P<0.05)。干扰素-γ释放试验和结核菌素皮内试验、T-SPOT TB和结核菌素皮内试验诊断肺结核的敏感性差异有统计学意义(P<0.05),特异性、阳性预测值、阴性预测值差异均无统计学意义(P>0.05)。干扰素-γ释放试验诊断肺结核的敏感性、特异性、阳性预测值和阴性预测值均稍高于T-SPOT TB试验,但差异均无统计学意义(P>0.05)。活动性肺结核组干扰素-γ释放试验阳性、阴性患者外周血白细胞计数、淋巴细胞绝对值、淋巴细胞百分比差异均无统计学意义(P>0.05)。结论化学发光法干扰素-γ释放试验方便、快捷,对辅助诊断肺结核有一定的价值。

薄壁空洞型肺结核和肺癌CT诊断与鉴别诊断

目的根据CT影像表现特征,结合γ干扰素释放试验(IGRA)及白细胞介素-6(IL-6),探讨薄壁空洞性肺结核与肺癌诊断与鉴别诊断价值。方法收集2016年2月-2020年2月临床及影像学资料完整的薄壁空洞型病灶73例,其中薄壁囊腔型肺癌组41例,薄壁空洞型肺结核组32例,均经术后病理确诊。对比分析两组病灶的的CT特征、IGRA及IL-6检测水平,并进行统计学分析。结果两组资料在性别、病灶大小及含气腔隙大小上差别不大,无统计学意义P>0.05;而在发病年龄、空洞壁厚较大差异,具有统计学意义P<0.05。结核组病灶的CT特点主要位于上叶尖后段及下叶背段,长毛刺、尖角征、单腔、腔内壁光整、卫星灶等发生率明显高于肺癌组,均有统计学意义P<0.05。肺癌组病灶的CT特点是边缘分叶、短毛刺、多腔、腔内壁不光整、内见分隔及血管穿行、磨玻璃征、蜂窝征等,均有统计学意义P<0.05。结核组IGRA阳性率、IL-6水平均明显高于肺癌组,均有统计学意义P<0.05。CT征象联合IGRA及IL-6,对薄壁空洞型肺结核与肺癌的诊断与鉴别诊断其灵敏度、特异度分别为1.000(41例)、0.875(28例)。结论分析薄壁空洞的CT特征,结合IGRA及IL-6水平,可以明显提高薄壁空洞型肺结核与肺癌的诊断与鉴别诊断,有利于临床制定治疗方案。

外周血中性粒细胞对γ干扰素释放试验结果的影响

目的探讨外周血中性粒细胞对γ干扰素释放试验(IGRA)阳性程度的影响。方法采集2020年10月—2022年3月上海市松江区中心医院234例临床诊断疑似肺结核患者全血样本,根据年龄将234例患者分为青少年组(<40岁)、中年组(40~<60岁)和老年组(>60岁)。检测所有患者血常规,提取外周血单个核细胞进行IGRA。收集所有患者常规检查结果和相关临床资料。将测试孔斑点数-空白对照孔斑点数11~14个(临近Cut-off值)定义IGRA弱阳性,>14个定义为IGRA阳性,<11个定义为IGRA阴性。分析不同年龄、性别患者中性粒细胞计数与IGRA阳性程度的关系。纳入2例临床病例进行验证。结果IGRA弱阳性患者中性粒细胞计数[(6.15±3.22)×10^(9)/L]显著高于阳性患者[(5.05±2.66)×10^(9)/L](P=0.0302);青少年组中,IGRA弱阳性患者中性粒细胞计数[(7.54±4.46)×10^(9)/L]显著高于阳性患者[(4.00±1.49)×10^(9)/L](P=0.0002);男性患者中,IGRA弱阳性患者中性粒细胞计数[(6.49±3.49)×10^(9)/L]高于阳性患者[(5.28±2.54)×10^(9)/L](P=0.0408)。临床病例验证结果显示,中性粒细胞计数低的患者更易出现IGRA阳性结果。结论中性粒细胞计数会影响IGRA阳性程度,建议在采用IGRA辅助诊断结核病时应排除中性粒细胞对IGRA结果的影响。

类紫杉醇脂质体体外释放的桨膜结合分析法研究

体外释放度是评价脂质体制剂质量的重要指标,目前各国药典中均未收载脂质体体外释放度评价方法,致使脂质体质量评价缺乏统一标准和无法提供安全性保障。本研究以自制类紫杉醇脂质体为例,通过优化外部释放条件建立的桨膜结合法模拟了在生理条件下类紫杉醇药物12 h的释放情况,结果表明以0.5%SDS-HEPES作为释放介质,采用截留分子量为1000 kD的透析袋对脂质体水溶液进行释放的结果满足要求,且具有区分能力,为载药脂质体体外释放方法开发提供参考。