基因芯片技术在矽肺合并分枝杆菌感染诊治中的应用

Clinical application of gene chip technology in diagnosis and treatment of silicosis complicated with mycobacterial infection

  • 摘要:
    背景 基因芯片法越来越广泛地用于普通结核的诊治,其在矽肺合并分枝杆菌感染诊治中的应用价值仍有待探索。
    目的 对基因芯片技术在矽肺合并分枝杆菌感染诊断及治疗中应用价值进行初步探讨。
    方法 以2019年1月—2021年6月经福建医科大学附属泉州第一医院诊治的197例疑诊合并分枝杆菌感染的矽肺患者为研究对象。197例患者分别采用痰涂片抗酸染色法(痰涂片法)、痰结核分枝杆菌培养法(痰培养法)、支气管肺泡灌洗液(BALF)基因芯片法3种方法进行分枝杆菌感染的病原学检测,其中80例患者在此基础上同时采用BALF涂片抗酸染色法(BALF涂片法)、BALF结核分枝杆菌培养法(BALF培养法)进行病原学检测。采用组内相关系数(ICC)比较各方法病原学检测阳性率的一致性。经BALF基因芯片法分枝杆菌菌种鉴定为结核分枝杆菌的标本,加做结核分枝杆菌耐药突变基因检测。
    结果 197例患者平均年龄为(53.1±9.1)岁,平均矽尘接触时间为(21.1±9.4)年,男性192例、女性5例;矽肺壹期8例、矽肺贰期17例、矽肺叁期172例;痰涂片法病原学检测阳性率为11.2%,痰培养法为24.4%,BALF基因芯片法为36.0%,组间差异均有统计学意义(均P<0.05)。一致性检验结果显示3种方法ICC为0.539(P<0.001)。80例患者中,5种方法病原学检测阳性率差异有统计学意义(χ2=25.23,P<0.001);两两比较结果显示,BALF培养法与痰涂片法、BALF涂片法,BALF基因芯片法与痰涂片法、BALF涂片法组间检测阳性率差异均有统计学意义(均P<0.05),其余组间差异无统计学意义(均P>0.05)。一致性检验结果显示5种方法ICC为0.586(P<0.001)。71例BALF基因芯片法检测阳性病例中:结核分枝杆菌复合群59例(17例检测出一线抗结核药的耐药基因,2例检测出二线抗结药喹诺酮的耐药基因),均接受规律抗结核治疗,其中45例病情好转,14例病情稳定;非结核分枝杆菌12例,5例接受抗非结核分枝杆菌治疗(其中4例病情好转,1例病情稳定),7例症状轻微,未接受抗非结核分枝杆菌治疗。
    结论 与痰涂片法、痰培养法等传统方法相比,BALF基因芯片技术可提高矽肺合并分枝杆菌感染病原学诊断阳性率,还能快速鉴别其是否为非结核分枝杆菌或耐药结核分枝杆菌感染,有助于临床尽早调整治疗方案。

     

    Abstract:
    Background Gene chip technology has been increasingly used in the diagnosis and treatment of common tuberculosis. However, its role in the diagnosis and treatment of silicosis complicated with mycobacterial infection remains unclear.
    Objective To evaluate the application value of gene chip technology in the diagnosis and treatment of silicosis complicated with mycobacterial infection.
    Methods From January 2019 to June 2021, 197 silicosis patients suspected to be complicated with mycobacterial infection in Quanzhou First Hospital Affiliated to Fujian Medical University were enrolled in this study. The etiology evaluation for the 197 patients was conducted by acid-fast staining of sputum smear (sputum smear method), culture of Mycobacterium tuberculosis of sputum (sputum culture method), and gene chip technology of bronchoalveolar lavage fluid (BALF); and for 80 patients among them, acid-fast staining of BALF (BALF smear method) and culture of Mycobacterium tuberculosis of BALF (BALF culture method) were additionally performed. The positive rates and consistency were assessed using intraclass correlation coefficient (ICC). Test for Mycobacterium tuberculosis drug resistance mutation gene was added for patients with Mycobacterium tuberculosis complex by BALF gene chip technology.
    Results The average age of the 197 patients was (53.1±9.1) years, and the average dust exposure time was (21.1±9.4) years, including 192 males and 5 females. There were 8 cases with stage I silicosis, 17 cases with stage II silicosis, and 172 cases with stage III silicosis. Among them, 11.2% were positive for sputum smear; 24.4% were positive for sputum culture, and 36.0% were positive by gene chip of BALF. The difference between the three methods was statistically significant (P<0.05). The result of consistency test for the three methods showed that the ICC was 0.539 (P<0.001). Among the 80 patients, there was a significant difference in the positive rates of the five methods (χ2=25.23, P<0.001). The results of Bonferroni test showed statistically significant pair-wise differences between BALF culture method and sputum smear method, BALF culture method and BALF smear method, BALF gene chip method and sputum smear method, BALF gene chip method and BALF smear method (P<0.05), while there were no statistically significant differences between the other pairs (P>0.05). The result of consistency test for the five methods showed that the ICC was 0.586 (P<0.001). Among the 71 BALF gene chip positive cases, 59 cases reported positive Mycobacterium tuberculosis complex (17 cases were positive in the first-line anti-tuberculosis resistance test, and 2 cases were found positive quinolone resistance gene in the second-line anti-tuberculosis resistance test), and received regular anti-tuberculosis treatment, among them 45 cases improved and 14 cases were stable; 12 cases reported non-tuberculous mycobacteria cases, among them 5 cases received anti-non-tuberculous mycobacteria treatment (4 cases improved and 1 case was stable), and 7 cases with mild symptoms did not receive anti-non-tuberculous mycobacteria treatment.
    Conclusion Compared with sputum smear, sputum culture, and other traditional methods, gene chip technology of BALF can improve the positive rate of pathogenic diagnosis of silicosis complicated with mycobacterial infection, and can also quickly identify whether it is non-tuberculous mycobacteria infection or drug-resistant Mycobacterium tuberculosis infection, which is helpful to adjust treatment as soon as possible.

     

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