Investigation of the cell composition and gene expression in the delayed-type hypersensitivity tuberculin skin test

Dear Editor,The tuberculin skin test(TST)reagents have continuously improved,with the ESAT6-CFP10(EC)test having recently been introduced,but are seldom based on the direction of the delayed-type hypersensitivity(DTH)mechanism.Previous studies only partially showed the infiltration and activation of immune cells and the production of cytokines of the skin induration[1,2],and lack the detailed measurements of cell proportions and gene expression in the DTH response.Therefore,in this study,we revealed the comprehensive characteristics of DTH by single-cell RNA sequencing(scRNA-seq)in the guinea pig tuberculosis(TB)model[Experimental Animal Welfare Ethics Committee,Beijing Tuberculosis and Thoracic Tumor Research Institute(2021-064)].

Performance and correlation of QuantiFERON-TB Gold, T-SPOT.TB and tuberculin skin test in young children with or exposed to tuberculosis

Objective:To evaluate the performance of interferon gamma release assays and tuberculin skin test in Bacillus Calmette-Guerin vaccinated young children.Methods:A cross-sectional study was conducted in healthy children younger than 5 years who were recently diagnosed with tuberculosis or had recent exposure to active tuberculosis.QuantiFERON-TB Gold,T-SPOT.TB and tuberculin skin test were performed in each patient.Results:Of the 60 children,median age 3.3 years,17 had tuberculosis and 43 had recent tuberculosis exposure.Overall,15(25.0%)children had tuberculin skin test reaction≥10 mm;8(13.3%)were positive by QuantiFERON-TB Gold In-Tube test,and 12(20.0%)by T-SPOT.TB.Nineteen(31.7%)children had at least one positive test.There was a moderate agreement between interferon gamma release assays and tuberculin skin test.Conclusions:The positive rates of interferon gamma release assays and tuberculin skin test were low in young children who were infected with tuberculosis,supporting the management strategy of not testing children younger than 5 years.

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.

Comparison between Quanti-FERON-TB Gold In-Tube test and tuberculin skin test for diagnosis of latent tuberculosis in children: A cross-section study

Objective: To compare Quanti-FERON-TB Gold In-Tube (QFT-GIT) test and tuberculin skin test (TST) for the diagnosis of latent tuberculosis infection in children. Methods: In this cross-sectional study, 64 participants who were between 3 months and 14 years old and had close contact with smear-positive pulmonary tuberculosis were included. Both QFT-GIT test and TST were done and the results were analyzed by SPSS software and Kappa test. Results: The distribution of gender and age according to QFT-GIT and TST results were matched (P>0.05). Overall agreement between QFT-GIT and TST for diagnosis of latent tuberculosis infection in children was 75%. In addition, the contingency coefficient was 0.257, and the Kappa measure of agreement was 0.246 (P=0.034). Conclusions: Compared to TST, QFT-GIT shows no apparent advantage for diagnosis of latent tuberculosis infection in children.

Indications, Interpretation and Clinical Consequences of Tuberculin Skin Tests in Resource Limited Settings

Objective: to evaluate the policy of TST testing in Suriname. As there is no gold standard to diagnose latent tuberculosis infection (LTBI), the tuberculin skin test (TST) is used to diagnose LTBI. However, internationally, the cut-off values of the TST are not uniform and depend on local tuberculosis (TB) epidemiology and guidelines for test initiation. In Suriname, where currently several indications exist for TSTs, cut-off values are set at 5 mm or 10 mm, depending on the age and/or medical history of the patient. LTBI classification is performed by pulmonologists primarily based on the American Thoracic Society targeted TB testing guidelines. Method: retrospective analysis of outpatient TST data between 2011 and 2019 from Suriname’s sole pulmonary medicine clinic. Result: 1373 patients were evaluated. 590 patients were from the screening group of whom 253 had a positive TST result, 46 of whom were classified as LTBI. In the contact tracing group of 649 patients, 616 had a positive TST, 352 of whom were classified as LTBI. In the medical condition group of 134 patients, 96 had a positive TST, 38 of whom were classified as LTBI. Eventually, positive TST results were found for 965 tested patients: 436 patients were classified as LTBI and 529 non-LTBI patients were not prescribed chemoprophylaxis. None of the non-LTBI TST-positive patients were diagnosed with active TB, including 174 patients with a TST result of 15 mm or greater and in need of IPT, but not prescribed by judgement of the pulmonologist or because of loss to follow-up. Conclusion: the overrepresentation of positive TST results in Suriname is attributable to stringent cut-off values, especially among patients who do not disclose TB risk factors. In our opinion the TST cut-off value for such patients in Suriname and other similar settings could be set at 15 mm. We also promote that for all patients with a TST result of 15 mm or greater, offering IPT should be considered (after excluding active TB).

BCG—PPD与TB—PPD皮肤试验在大学生结核病筛查中的差异性分析

目的 比较卡介菌纯蛋白衍生物（purified protein derivative of BCG，BCG-PPD）与结核菌素纯蛋白衍生物（purified protein derivative of tuberculin,TB-PPD）在大学生结核病筛查中的差异。 方法 回顾性分析2012—2013年北京市西城区3所大学2821名学生的结核病筛查数据。根据使用的PPD试剂种类进行分组，BCG-PPD组学生1386名；TB-PPD组学生1435名。对两组观察者分别进行结核菌素试验，72 h查验硬结平均直径，对强阳性者进行胸部X线检查。采用SPSS 11.5软件进行统计分析，用秩和检验对两组硬结平均直径进行比较,用χ^2检验对组间PPD结果分布频度及结核病筛查结果进行比较，当理论值＜1时，采用Fisher确切概率法，卡痕与PPD阳性结果间相关性分析采用logistic回归，P〈0.05为差异有统计学意义。结果 中位硬结平均直径BCG-PPD组（6 mm）大于TB-PPD组（0 mm），差异有统计学意义（Z=－10.034, P〈0.05）。BCG-PPD组的阳性率（54.8%, 759/1386）明显高于TB-PPD组（32.7%,469/1435）（χ^2=139.818，P〈0.05）；PPD强阳性率BCG-PPD组为6.6%（92/1386）,高于TB-PPD组的3.4%（49/1435）（χ^2=15.425，P〈0.05）；BCG-PPD组的异常反应发生率为1.9%（27/1386），高于TB-PPD组的0.4%（6/1435），差异有统计学意义（χ^2=14.274，P〈0.05）。卡痕与PPD阳性结果相关，有卡痕者更容易出现PPD阳性结果（OR=2.046，95％CI=1.630～2.569，P〈0.05）。 结论 TB-PPD和BCG-PPD皮肤试验在大学生结核病筛查中的强阳性率和异常反应发生率上存在差别，有进一步探讨的必要。

脐血lgE水平、PPD皮试反应与小儿过敏性疾病

为了观察脐血IgE含量、不同年龄小儿PPD皮试反应与小儿常见过敏性疾病的关系 ,探讨Th1、Th2应答失常与过敏性疾病的关联 ,进一步了解过敏性疾病的免疫发病机理 ,采用ELISA法检测204例脐血血清IgE水平 ,随访2岁龄幼儿湿疹、过敏性鼻炎、哮喘性支气管炎的发生情况 ,同时调查2岁龄幼儿和7岁龄小学生PPD皮试反应与常见过敏性疾病发生情况。结果表明 :①脐血IgE增高小儿有过敏性鼻炎、哮喘性支气管炎高发倾向。②2岁龄幼儿过敏性鼻炎、哮喘性支气管炎总的人次以PPD( -)组为多 ;有家族过敏史、反复呼吸道感染的PPD( -)年长儿患过敏性疾病机率高于PPD(+)者。提示 :脐血IgE。

T-SPOT.TB与PPD在消化系统结核感染鉴别诊断中的价值

目的评估快速T细胞计数体外酶联免疫斑点技术(T-ELISPOT.tuberculosis,T-SPOT.TB)与PPD试验在消化系统结核感染的诊断价值。方法回顾性分析2011年6月-2013年7月于南京军区南京总医院消化内科收治的60例患者,其中14例确诊为结核感染的患者,46例为非结核感染的消化系统疾病患者,抽取患者外周血送检,T-SPOT.TB与PPD试验结果的准确性进行比较。结果 14例确诊结核感染患者,46例非结核感染的患者,T-SPOT.TB的灵敏度与PPD试验相近,差异无统计学意义(78.6%vs71.4%,P=1.000);T-SPOT.TB的特异性显著大于PPD试验(91.3%vs 69.6%,P=0.009);T-SPOT.TB阳性预测率显著大于PPD试验(73.3%vs 41.7%,P=0.001);T-SPOT.TB阴性预测率与PPD试验相近,差异无统计学意义(93.3.%vs 91.1%,P=1.000)。T-SPOT.TB与PPD试验在诊断消化系统结核感染方面有较好的吻合性(K=0.341,P=0.018)。结论 T-SPOT.TB应用于消化系统感染的诊断,尤其适合PPD阳性的疑似消化系统结核感染患者的鉴别诊断。

肺结核病人PPD皮试与ELISPOT结果的相关性分析

[目的]研究肺结核病人酶联免疫斑点技术(ELISPOT)和结核菌素纯蛋白衍生物(PPD)皮试结果的相关性。[方法]24例痰结核菌培养阳性肺结核病人,38例临床疑似肺结核病人,取外周血分离获取单个核细胞(PBMCs)后行ELISPOT试验。[结果]痰结核菌培养阳性肺结核组与临床疑似肺结核组PPD皮试阳性率相似;痰结核菌培养阳性肺结核组的ELISPOT阳性率高于临床疑似肺结核组;ELISPOT斑点数与PPD皮试反应程度之间缺乏相关性。[结论]ELIS-POT的诊断特异性高于PPD皮试。

北京市怀柔区某高校大学入学新生PPD皮肤反应特征分析

目的分析北京市怀柔区某高校入学新生结核菌素纯蛋白衍生物(TB-PPD)皮肤试验的阳性水平,为高校结核病防控工作提供参考性建议。方法以怀柔区某高校2014-2019学年入学的4048名新生为研究对象,通过对新生结核病筛查数据回顾性研究,分析该高校新生自然感染状态下TB-PPD皮肤试验基线水平及影响因素。结果2014-2019年怀柔区某高校入学新生PPD皮肤试验阳性率、强阳性率呈下降趋势。全部入学新生TB-PPD皮肤试验阳性率为16.38%,强阳性率为2.67%。其中男、女生强阳性率分别为2.87%和2.55%,性别间差异无统计学意义(χ^(2)=0.369,P=0.544)。按新生生源地省份划分为东、中、西部三组,χ^(2)检验显示三组间TB-PPD皮肤试验强阳性率的差异无统计学意义(χ^(2)=0.869,P=0.647)。PPD皮肤试验强阳性率随新生入学年龄升高而增加(r=0.927,P=0.023)。结论该高校TB-PPD皮肤试验强阳性率与入学年份及入学年龄相关,建议学校了解本地高校新生PPD皮肤试验反应率水平及趋势,重点关注高年龄入学新生的PPD结果,当群体反应率水平有较大波动或招收新生整体年龄上升时要做好应对风险准备。

PPD皮试在继发性肺结核诊断中意义的分析

目的通过对国内有关继发性肺结核与PPD皮试联系的病例对照研究结果进行荟萃分析（meta analysis），获得综合的结论，以期对PPD皮试作为诊断实验在诊断继发性肺结核中的意义作出评价。方法以“PPD”为关键词，选用中国学术期刊全文数据库（cHKD），检索1994—2006年国内已发表的相关文献，选择符合标准的文献进行荟萃分析。对所有研究予以加权定量合并，计算灵敏度和特异度及其95％可信区间。绘制汇总受试者工作特征曲线（SROC），并计算曲线下面积。结果共纳入8篇中文文献，研究对象共计1479人，按照随机效应模型计算汇总数据，得优势比为exp（A）=e）cp（2．2998）=9．9726，可以认为异常组阳性优势明显高于正常组。诊断试验的准确度指标TPR＊=0．7595，其标准误亚（TPR＊）=0．04915。结论汇总目前关于PPD皮试诊断继发性肺结核的研究显示，PPD皮试是一种准确度较高的检查方法，但还需要高质量的前瞻性研究以更准确地评价其临床价值。

结核菌素纯蛋白衍生物(TB-PPD)临床安全性的研究

目的评价结核菌素纯蛋白衍生物(TB-PPD)的临床安全性。方法随机选取50例健康志愿者,对其皮内注射0.1mL TB-PPD试剂,注射结束后分别于30min、24h、48h、72h和7d观察其不良反应发生率,评价其安全性。结果不良反应主要表现为发热、接种部位疼痛、硬结等,未发现严重不良反应。结论 TB-PPD具有较好的临床安全性。

分析某高校新生PPD结果及意义

[目的]探讨PPD结果与肺结核发病的关系,为制定肺结核的预防措施提供依据。[方法]采用PPD0.1ml(5IU)在左前臂屈侧作皮内注射,72h测量皮肤硬结直径,根据硬结直径情况分为一般阳性、强阳性两组,对阳性者拍摄胸部正位片、验血、查痰等检查。[结果]13446名新生中,PPD阳性1743名,一般阳性率为9.3%,强阳性率为3.7%,PPD阳性者65例胸片异常,强阳性者5例确诊活动性肺结核,而PPD一般阳性者无一例出现肺结核。[结论]PPD检查是一种简单易行的筛检肺结核的方法,根据试验结果采取不同防治措施,可有效控制结核病在高校的传播和流行。

采用干扰素释放反应试验和PPD皮试对深圳市高校学生结核分枝杆菌潜伏感染筛查的研究

目的采用结核分枝杆菌特异性IFN-γ免疫酶联斑点法(ELISPOT)和传统的结核菌素皮试(PPD皮试)对深圳市高校学生结核分枝杆菌潜伏感染现状进行筛查,并比较两种方法的差异。方法采用自主研制的Elispot试剂盒(简称Elispot)和PPD皮试,对133例健康志愿者进行平行检测。结果A高校99例志愿者中PPD皮试有49例阳性(49.5%),Elispot检测有20例阳性(20.2%);B高校34例志愿者中PPD皮试有27例阳性(79.4%),Elispot检测有2例阳性(5.9%)。对两种方法进行平行比较分析,结果有统计学差异(χ2=41.31,P<0.005);在PPD()组中Elispot阳性率最高为28.57%;在()-(-)组中分别为22.2%、12.5%及12.28%。结论结核菌特异性IFN-γElispot试剂盒在健康人群中检测阳性率比传统的PPD皮试要低,在诊断结核分枝杆菌潜伏感染的价值优于PPD皮试,在结核分枝杆菌潜伏感染早期诊断有较好的发展前景。

常规人型-PPD、BCG-PPD、PHA三联皮肤试验的临床应用及评价

对1097例门诊和住院病人进行常规人型-PPD、BCG-PPD、PHA三联皮肤试验。结果表,患病人群中PPD强阳性率随年龄增长而增高,高峰年龄30～50岁,患病人群平均PPD强阳性率为26.07％～31.09％;PHA阳性率为79.58％;临床疑诊结核病、购膜炎、消化道疾患等PPD阳性率较高;12岁以下小儿一般阳性率显著低于全国卡介苗保护率平均水平。常规PPD检测不仅对结核病早期诊断、细胞免疫功能评价有重要意义,而且在评价卡介苗接种效果、及时发现漏种病例、提高整体免疫水平方面亦有重要价值。

1000例卡介苗接种儿童BCG-PPD皮试反应的结果分析

目的 :了解 5 14例卡介苗接种后的健康儿童和 486例伴发有感染性疾病的儿童进行 BCG- PPD皮试反应的结果。方法 :用 BCG- PPD皮试法检测阳转率。结果 :健康儿童 5 IU的转阳率为 70 % ,疾病儿童为 5 5 %。健康儿童的转阳率明显高于感染儿童。 BCG- PPD皮试后硬结直径 ,健康儿童与疾病儿童比较无统计学差异。结论 :感染疾病可影响皮试的转阳率 ,对 BCG (卡介苗 )接种后儿童的监测建议尽量避免儿童在感染时期进行 BCG- PPD皮试 。

T-SPOT.TB测定、PPD试验和结核蛋白芯片对儿童肺结核诊断价值的研究

目的利用T-SPOT.TB测定、结核菌素试验方法(PPD法)及结核蛋白芯片法进行儿童结核分枝杆菌的检测诊断比较,以评价各种不同方法在儿童肺结核诊断中的临床应用价值。方法回顾性分析2015~2017年宜昌市第三人民医院收治的疑似肺结核患儿108例,其中确诊肺结核患儿57例,非结核患儿51例,用上述3种方法分别检测108例患儿,统计分析三种方法在诊断儿童结核病的灵敏度、特异度,并比较T-SPOT.TB与PPD法及结核蛋白芯片法诊断儿童肺结核的准确度。结果 T-SPOT.TB测定法、PPD法及结核蛋白芯片法检测儿童肺结核的灵敏度分别为87.72%、56.14%和52.63%;特异度分别为90.19%、66.67%和82.35%。结论 T-SPOT.TB测定与PPD法及结核蛋白芯片法比较,具有快速、敏感性高和特异性强等优点,是临床儿童结核病快速诊断的有效检测方法。

PPD皮试联合结核感染T淋巴细胞斑点试验检测对肺结核的诊断价值

目的探究在肺结核的诊断中PPD皮试联合结核感染T淋巴细胞斑点试验检测的诊断价值。方法选择2019年1月至2020年8月期间在我院接受治疗的100例肺结核患者作为临床研究对象为观察组,并选取100例同期在我院接受治疗的非肺结核性肺部疾病患者做对比对象为对照组,所有患者均接受PPD皮试以及结核感染T淋巴细胞斑点试验检测。分析在肺结核的诊断中PPD皮试联合结核感染T淋巴细胞斑点试验检测的诊断价值。结果检测发现,观察组患者的PPD皮试阳性率(47.00%)以及结核感染T淋巴细胞斑点试验检测结果阳性率(85.00%)显著高于对照组患者的PPD皮试阳性率(30.00%)以及结核感染T淋巴细胞斑点试验检测结果阳性率(1.00%),差异具有统计学意义(P<0.05);观察组患者中,联合检测准确率(98.00%)显著高于PPD皮试诊断准确率(47.00%)以及结核感染T淋巴细胞斑点试验检测准确率(85.00%),差异具有统计学意义(P<0.05)。结论在肺结核的诊断中PPD皮试联合结核感染T淋巴细胞斑点试验检测具有显著临床价值,诊断准确率更高。

Analysis of the chemoprophylactic effect on close contacts of patients with active tuberculosis and positive tuberculin skin tests

Objective:The current study analyzed the chemoprophylactic effect of isoniazide on close contacts of patients with active tuberculosis and positive tuberculin skin tests(TSTs).Methods:A total of 1206 close contacts of patients with active tuberculosis and strongly-pos-itive TSTs were enrolled.The patients had chest X-ray examinations.Patients without tuberculosis and other diseases were divided into the following groups:90 patients in the prophylaxis group,who were given 300 mg of isoniazid qd(3-5 mg/kg for children)over a 10-month treatment course;and 89 patients in the control group without drug therapy.Both groups were followed for 10 years.Results:(1)There were 568 patients with negative results and 638 with positive results,includ-ing 445 with ordinarily-and moderately-positive results,and 193 with strongly-positive results(a positive rate of 52.9%[638/1206]and a strongly-positive rate of 16.0%[193/1206]).Fourteen tuber-culosis patients were identified(tuberculosis detection rate of 1.1%).(2)During the 3-year period of follow-up,there were 4 patients in the prophylaxis group and 12 patients in the control group who acquired tuberculosis(a morbidity rate of 4.7%[4/84]and 13.4%[12/89],respectively),and the dif-ference was statistically significant(χ^(2)=3.916,P=0.048).Six patients in the prophylaxis group,all of whom were children,discontinued medication use during the course of treatment due to adverse drug reactions,for an adverse drug reaction occurrence rate of 6.6%(6/90),a medication completion rate of 93.3%(84/90),and a 3-year protection ratio of 64.6%.(3)During the 4-6 year period,there were two patients in the prophylaxis group and three patients in the control group who acquired tubercu-losis(a morbidity rate of 2.5%[2/78]and 4.1%[3/73],respectively),two in the prophylaxis group and four in the control group who were lost to follow-up(a loss to follow-up rate of 2.5%[2/80]and 5.1%[4/77],respectively),and the difference was not statistically significant(χ^(2)=0.006,P=0.940;χ^(2)=0.215,P=0.643).(4)During the 7-10 year study,there was one patient in the prophylaxis group and two patients in the control group who acquired tuberculosis(a morbidity rate of 1.3%[1/72]and 3.1%[2/64],respectively),and four in the prophylaxis group and six in the control group who were lost to follow-up(a loss to follow-up rate of 5.2%[4/76]and 8.5%[6/70],respectively),and the dif-ference was not statistically different(χ^(2)=0.011,P=0.918;χ^(2)=0.176,P=0.675).(5)Within 10 years,there were 7 patients in the prophylaxis group and 17 patients in the control group who acquired tu-berculosis(a morbidity rate of 8.3%[7/84]and 21.5%[17/79],respectively;χ^(2)=4.770,P=0.029),and 6 in the prophylaxis group and 10 in the control group were lost to follow-up(the loss to follow-up rate was 7.1%[6/84]and 11.2%[10/89],respectively;χ^(2)=0.863,P=0.353).Conclusion:Close contacts of patients with active tuberculosis are at high-risk for acquir-ing tuberculosis.It is safe and effective for patients with strongly-positive TST results to undergo isoniazid chemoprophylaxis for 10 months.

基于改进YOLOv5的人体结核菌素试验后皮下硬块检测算法研究

针对传统皮下硬块检测方法精度低、计算速度慢等问题,提出了一种基于改进YOLOv5的皮下硬块目标检测算法。首先,将YOLOv5模型的主干网络CSPDarknet53进行改进,引入Faster模块来替换其中的C3模块。其次,利用模型剪枝在保证整个模型性能的同时减轻其计算复杂度。最后,引入Wise-IoU来进一步提升网络的回归性能。实验结果表明,基于改进YOLOv5的皮下硬块目标检测算法相比于原始的YOLOv5,准确率提升了1.2%,参数量减少了77.7%,整个算法更加轻量化,有效提高了算法对于皮下硬块的检测精度,减少了计算参数量,提升了算法的运行速度。