支气管肺泡灌洗液涂片抗酸染色、L-J培养、TB-LAMP、Xpert MTB/RIF检测在肺结核诊断中的应用比较

目的比较支气管肺泡灌洗液(BALF)涂片抗酸染色、罗氏固体培养(简称"L-J培养")、环介导恒温扩增技术(TB-LAMP)、结核分枝杆菌(MTB)多色巢式荧光定量PCR(Xpert MTB/RIF)检测在肺结核诊断中的应用价值。方法收集BALF样本165例份,165例患者中临床诊断为肺结核131例、其他呼吸道疾病34例。每例份样本同时进行涂片抗酸染色镜检、L-J培养、TB-LAMP和Xpert MTB/RIF检测。结果 165例份BALF样本中,抗酸染色、L-J培养、TB-LAMP和Xpert MTB/RIF检测阳性率分别为15.15%(25/165)、24.24%(40/165)、40.00%(66/165)、29.09%(48/165)。在抗酸染色抗酸杆菌(AFB)阳性样本中,L-J培养、TB-LAMP及Xpert MTB/RIF检测MTB阳性率分别为84.00%(21/25)、100.00%(25/25)、96.00%(24/25),3种方法检测MTB阳性率比较P均> 0.05;在抗酸染色AFB阴性样本中,L-J培养、TB-LAMP及Xpert MTB/RIF检测MTB阳性率分别为13.57%(19/140)、29.29%(41/140)、17.14%(24/140),两两比较P均<0.05。以临床诊断为金标准,TB-LAMP检测诊断肺结核的灵敏度高于L-J培养、Xpert MTB/RIF检测(P均<0.05);L-J培养、Xpert MTB/RIF检测诊断肺结核的灵敏度比较无统计学差异(P> 0.05);L-J培养、TB-LAMP及Xpert MTB/RIF检测诊断肺结核的特异度均为100%。结论 BALF涂片抗酸染色AFB阳性率最低,只适合肺结核的初筛;L-J培养和TB-LAMP、Xpert MTB/RIF检测均能大幅提高抗酸染色AFB阴性肺结核的病原学诊断率,其中TB-LAMP检测的阳性率及其诊断肺结核的灵敏度最高。

Clinical Performances of Pure TB-Lamp Kit for <i>M. tuberculosis</i>Complex Detection in Sputum Samples

Tuberculosis represents a main concern for public health worldwide. In poor countries, the most prevalent method for bacteriological confirmation re- mains Smear Sputum Microscopy (SSM). This study objective was to assess clinical performances of Loop Mediated Isothermal Amplification for TB detection (Lamp-TB). Sputum of patients presenting symptoms consistent with tuberculosis were collected according to the National Tuberculosis Control Programme guidelines in Centre Antituberculeux de Yopougon. SSM after Ziehl-Neelsen staining and TB-Lamp were blindly performed with spot sputum specimen. Samples, transported at Institut Pasteur de Cote d’Ivoire were decontaminated according to N-acetyl-L-cystein (NALC) method. In Mycobacteria Growth Indicator Tube (MGIT), 500 μl of pellet were inoculated and incubated in MGIT 960 instrument. MPT64 antigen was detected on positive culture. Of 500 patients enrolled, 469 were included. Clinical isolates of M. tuberculosis Complex were detected for 157 (33.5%). Comparatively to culture, Sensitivity and Specificity of SSM were 86% (95% Confidence interval (CI): 81% - 91%) 96% (95%IC: 94% - 98%) respectively. TB-Lamp Sensitivity was 92% (95%CI: 88% - 96%), and Specificity 94% (95%CI: 91% - 97%). Positive Predictive Value of SSM and TB-Lamp was 91.8% and 88.8% respectively. Negative Predictive Value of TB-Lamp assay was 95.7% whereas this of SSM was 93.3%. Positive Likelihood Ratio was 15.3 for TB-Lamp and 21.5 for SSM 21.5 whereas negative Likelihood of TB-Lamp was lower than SSM. Active tuberculosis was detected in162/469 (34.5%) with TB-Lamp and 147 (31.3%) with SSM. TB-Lamp assay performances estimated from sputum samples may improve detection of active TB cases in routine.

快速、简便、准确 LAMP~技术深耕医疗领域 中国LAMP~研究会第五届大会暨TB-LAMP新品发布会在大连召开

日前,由日本荣研化学株式会社、荣研生物科技（中国）有限公司主办,北京蓝谱生物科技有限公司与北京蓝谱生物技术开发有限公司协办的＂中国LAMP（R）研究会第五届大会暨TB-LAMP新品发布会＂在大连召开,一款可以在一个小时内完成结核杆菌检测的产品——TB-LAMP引起与会者的高度关注,进而引发业内对LAMP（R）技术最新研究成果的跟进,关于LAMP（R）技术在结核病诊断、肿瘤细胞检测、埃博拉病毒检测、水产品安全检验等方面的研究以及应用成果,成为此次会议讨论的热点。

WHO发表TB-LAMP在结核病检测中应用评价报告

结核病是引起全球死亡数前十位中死亡最高的疾病,仅仅是继艾滋病毒、艾滋病之后,是由单一传染性病原体引起的最大杀手,全世界已有近1/3的人口受到结核分支杆菌的感染,2013年,有900万人罹患结核病,150万人死于该疾病。95%以上的结核病死亡发生在低收入和中等收入国家,它还是导致15至44岁妇女死亡的五大原因之一,

TB-LAMP联合夹层杯集菌离心涂片法快速检测对提高尘肺合并结核阳性率的研究

目的探讨TB-LAMP联合夹层杯集菌离心涂片法快速检测对提高尘肺合并结核阳性率的应用价值。方法选取我院在2018年1月-2019年1月就诊的尘肺患者共150例作为研究对象,用直接涂片法、BD960培养法、TB-LAMP及夹层杯集菌离心涂片法进行检测,观察对比4种检验方法结核分枝杆菌的的阳性率。结果BD960培养法、TB-LAMP及夹层杯集菌离心涂片法对比直接涂片法的阳性率结果均差异有统计学意义,P<0.05,但是前三者之间阳性率结果没有明显差异;随着尘肺期别的提高,尘肺合并结核的比例也升高,结核分枝杆菌的阳性率也升高,BD 960培养法、TB-LAMP及夹层杯集菌离心涂片法对比直接涂片法在尘肺叁期及总数上的阳性率结果均差异有统计学意义,P<0.05,但是前三者之间的阳性率结果没有统计学意义;TB-LAMP联合夹层杯集菌离心涂片法快速检测和直接涂片法的阳性率比较差异有统计学意义(P<0.01)。结论TB-LAMP联合夹层杯集菌离心涂片法快速检测能明显提高尘肺合并结核的阳性率,具有广泛的应用价值。

以肺泡灌洗液行结核/非结核分枝杆菌检测对菌阴肺结核的诊断价值

目的评估取支气管肺泡灌洗液(BALF)进行结核/非结核分枝杆菌(MTB/NTM)检测对菌阴肺结核的诊断价值。方法收集临床诊断为菌阴肺结核的324例患者资料,对上述患者进行支气管镜检查,所有患者取BALF标本,根据检测方法分为MTB/NTM组(进行罗氏培养、涂片抗酸染色、MTB/NTB检测)、利福平耐药实时荧光定量核酸扩增检测技术(GeneXpert MTB/RIF)组(进行罗氏培养、涂片抗酸染色、GeneXpert MTB/RIF检测)、结核分枝杆菌环介导等温扩增技术(TBLAMP)组(进行罗氏培养、涂片抗酸染色、TB-LAMP检测)。结果MTB/NTM组罗氏培养、涂片抗酸染色、MTB/NTM检测阳性率分别为14.04%、1.75%、43.86%;GeneXpert MTB/RIF组罗氏培养、涂片抗酸染色、GeneXpert MTB/RIF检测阳性率分别为16.77%、3.73%、54.66%;TB-LAMP组罗氏培养、涂片抗酸染色、TB-LAMP检测阳性率分别为18.87%、4.72%、49.06%。MTB/NTM、GeneXpert MTB/RIF、TB-LAMP检测分别与罗氏培养、涂片抗酸染色阳性率比较,差异均有统计学意义(P<0.05)。MTB/NTM检测与GeneXpert MTB/RIF检测、TB-LAMP检测比较,差异无统计学意义(χ^(2)=2.185,P=0.335)。结论MTB/NTM检测具有较好诊断效能,相对简单、快捷,相较于TB-LAMP检测、GeneXpert检测更经济,并能鉴别NTM,及时减少NTM病患者误诊后使用结核药引起身体损害及经济损失。

铝酸盐绿色荧光粉制备工艺与发光性能关系的研究

采用高温固相法制备了CeMgAl11O19∶Tb(CMAT)荧光粉,系统研究了该荧光粉的制备工艺与发光性能的关系,并对不同结果进行了分析。研究表明,铝含量适当过量有利于荧光粉发光亮度的提高;在1400～1550℃的范围内,CMAT的发光亮度呈稳定的上升趋势,这表明在一定的范围内,温度越高越有利于材料的结晶;MgF2可以取代目前工业生产中普遍采用的传统助熔剂H3BO3,起到很好的激活晶格形成,促进晶粒生长的效果。

均相荧光探针PcR技术在TB检测中的研究

本实验利用均相荧光探针 Pc R技术对结核病患者痰液进行了检测 ,结果说明 ,采用均相荧光测量方式 ,提高了 Pc R检测的特异性与灵敏度 ,同时使实验操作步骤简化 ,杜绝扩增产物污染的途径及人工观察电泳的主观性 ,可适用于各种病原体、点突变等基因诊断 ,使 Pc

环介导等温扩增技术用于结核病诊断的价值评估

目的评估环介导等温扩增技术(loop-mediated isothermal amplification,LAMP)在诊断结核病中的应用价值。方法以疑似活动性结核患者为研究对象。前瞻性收集患者临床标本,分装后进行荧光涂片显微镜镜检、BACTEC MGIT 960液体培养、MTB GeneXpert(Xpert)和LAMP检测,分析比较这4种检测方法的灵敏度和特异度,同时进行一致性检验。结果研究共入组315例患者,分为痰标本组和非痰标本组,两组分别有175例和80例纳入分析。对于痰标本的检测,以液体培养为诊断标准时,LAMP的灵敏度为78.9%,特异度为82.7%;在有抗结核活性药物暴露史亚组中,LAMP的灵敏度为92.3%,Xpert的灵敏度为88.5%;以临床诊断为诊断标准时,LAMP的灵敏度优于涂片(58.9%vs.46.0%,P<0.05),特异度为98.0%;亚组分析提示在有抗结核活性药物暴露史的患者标本中,LAMP的灵敏度高于其他检测方法。对于非痰标本的检测,同样显示出优于涂片的灵敏度。结论LAMP在结核病的诊断中具有较好的灵敏度和特异度,与Xpert检测结果一致性较高,与培养一致性中等,优于结核菌涂片法。在有抗结核活性药物暴露史的患者中,LAMP有可靠的检出率。

Performance of Loop-Mediated Isothermal Amplification (LAMP)-Lateral Flow Detection and Xpert MTB/RIF Ultra for Diagnosis of Pulmonary Tuberculosis

Background: Due to the limitations of diagnosis by Xpert MTB/RIF assay, WHO suggests Loop—Mediated Isothermal Amplification (TB-LAMP) instead of sputum-smear microscopy for pulmonary TB diagnosis in patients. Dr. Thongchai Kaewphinit et al. invented a fast TB detection kit called TB d-tect (LAMP-LFD assay). There was no clinical trial to estimate the performance of TB d-tect. Objective: This study was aimed to find the performance of LFD assay and Xpert MTB/RIF ultra for pulmonary TB. Material and methods: A cross-sectional study was conducted. Suggestive pulmonary TB patients were enrolled from June 2020-28 February 2021. Respiratory specimens were collected from each patient and sent for AFB smear, LAMP-LFD assay, Xpert MTB/RIF ultra and culture TB. Result: 139 patients with suspected pulmonary TB were enrolled. 51% of patients were diagnosed with pulmonary tuberculosis. Based on culture TB as a gold standard, the sensitivity and specificity of LAMP-LFD assay were 85.4% (95% CI: 70.8% - 94.4%) and 87.8% (95% CI: 79.6 - 93.5), respectively. The sensitivity and specificity of Xpert MTB/RIF ultra were 95.1% (95% CI: 83.5% - 99.4%) and 74.5% (95% CI: 64.7 - 82.8), respectively. According to ROC curve, it was found that the areas under the curve of LAMP-LFD assay and Xpert MTB/RIF ultra were 0.866 and 0.848, respectively (p = 0.546). Conclusion: The diagnostic sensitivity and specificity of LAMP-LFD assay appeared to be comparable to those of Xpert MTB/RIF ultra. LAMP-LFD assay could be used in resource limiting laboratory.