Mutation Characteristics of inhA and katG Genes in Isoniazid-Resistant Mycobacterium Tuberculosis Patients in Xinjiang

Objective:To analyze the mutation characteristics of inhA and katG genes in isoniazid-resistant Mycobacterium tuberculosis in Xinjiang.Methods:The katG and inhA in 148 strains of isoniazid-resistant Mycobacterium tuberculosis were amplified through fluorescence quantitative PCR,and the amplified products were sequenced and compared.Results:The inhA gene mutation rate of 148 strains of isoniazid-resistant mycobacterium tuberculosis was 13.51%(20/148),among which the inhA gene mutation rate among patients of Han,Uygur,and Kazakh ethnicity were 15.87%,13.21%,and 17.65%,respectively.There was no significant difference in the inhA mutation rate among nationalities(c^(2)=2.897,P>0.05).The mutation rate of the katG gene was 84.46%(125/148),among which the mutation rates of patients of Han,Uyghur,and Kazak ethnicities were 82.54%,84.91%,and 76.47%,respectively.The Hui and other ethnic groups were all affected by the katG gene mutation.There was no significant difference in the mutation rate of the katG gene among different ethnicities(c^(2)=3.772,P>0.05).The mutation rates of the inhA gene in southern Xinjiang,northern Xinjiang,and other provinces were 18.60%,9.28%,and 37.50%,respectively.The mutation rates of the inhA gene in different regions were statistically different(c^(2)=6.381,P<0.05).There was no significant difference in the inhA mutation rate between patients from southern and northern Xinjiang(c^(2)=2.214,P>0.05)and between southern Xinjiang and other provinces(c^(2)=1.424,P>0.05).However,the mutation rate of the inhA gene in patients from other provinces was higher than that in northern Xinjiang(c^(2)=5.539,P<0.05).The mutation rates of the katG gene in southern Xinjiang,northern Xinjiang,and other provinces were 81.40%,87.63%,and 62.50%,respectively.There was no significant difference in the mutation rates of the katG gene among different regions(c^(2)=3.989,P>0.05).Conclusion:katG gene mutation was predominant in isoniazid-resistant tuberculosis patients in Xinjiang Uygur Autonomous Region,and inhA and katG gene mutation were no different among different ethnic groups.

Involvement of 2'-5'oligoadenylate synthetase-like proteinin the survivalof Mycobacterium tuberculosis avirulent strain in macrophages

Mycobacterium tuberculosis(M.tuberculosis)can replicate in the macrophage by interfering with many host protein functions.While it is far from known these host proteins for controlling M.tuberculosis infection.Herein,we infected macrophages including THP-1 and Raw264.7 cells with M.tuberculosis and identified the differentially expressed genes(DEGs)in the interferon signaling pathway.Among them,2'-5'oligoadenylate synthetase-like(OASL)underwent the greatest upregulation in M.tuberculosis-infected macrophages.Knockdown of the expression of OASL attenuated M.tuberculosis survival in macrophages.Further,bioinformatics analysis revealed the potential interaction axis of OASL-TAB3-RvO127,which was further validated by the yeast-two-hybrid(Y2H)assay and Co-IP.This interaction axis might regulate the M.tuberculosis survival and proliferation in macrophages.The study reveals a possible role of OASL during M.tuberculosis infection as a target to control its propagation.

Prokaryotic expression,identification and bioinformatics analysis of fbpB-esxA fusing gene from Mycobacterium tuberculosis

Objective:To obtain fbpB-esxA fusing gene of Mycobacterium tuberculosis（MTU）,express the encoded fusing protein in Escherichia coli（E.coli）,identify protein acquired,and predict the structure and function of the protein utilizing methods of bioinformatics.Methods:fbpB and esxA gene were amplified from genome of MTB H37Rv by PCR.The fbpB-esxA fusing gene Iigated by（Gly4Ser）3 linker was gained by means of Gene Splicing by Overlapping Extension PCU（SOEPCR）, and fusing gene was cloned into expression vector pET-30a.The recombinant plasmid was sequenced and expressed in E.coli BL21（DE3）.The protein was identified by Western blot using anti-HIS antibody.Secondary structure and antigenic epitopes of the protein were predicting using tools of bioinformatics.Results:The UNA sequences fbpB-esxA were identical with that published by GenBank.The Ag85B-ESAT-6 fusion protein about 50 kDa comprised 485 amino acids was efficiently produced from expression system in E.coli B1.21（DE3） under the induction of IPTG.Bioinformatics analysis showed the protein contained one transmembrane region and fourteen potential antigenic epitopes.Conclusions:The Ag85B-ESAT-6 fusion protein is successfully expressed with N-terminal HIS-tag.Gel filtration demonstrated that it exists as insoluble inclusion bodies mainly.The existence of linker doesn’t affect immunogenicity of Ag85B and ESAT-6.It will allow lor characterization in vitro and establish a foundation of further function research such as vaccine or diagnostic reagent.

Construction, Expression and Identification of a Recombinant BCG Vaccine Encoding Human Mycobacterium Tuberculosis Heat Shock Protein 65

Heat shock protein 65 (HSP65) is one of the most important protective immunogens against the tuberculosis infection. The signal sequence of antigen 85B and the whole HSP65 DNA sequence of human Mycobacterium tuberculosis (M. tuberculosis) were amplified from BCG genome and plasmid pCMV-MTHSP65 respectively by polymerase chain reactions (PCR). These two sequences were cloned into the plasmid pBCG-2100 under the control of the promoter of heat shock protein 70 (HSP70) from human M. tuberculosis, yielding the prokaryotic shuttle expression plasmid pBCG-SP-HSP65. Results of restriction endonuclease analysis, PCR detection and DNA sequencing analysis showed that the two cloned DNA sequences were consistent with those previously reported, and the direction of their inserting into the recombinant was correct and the reading frame had been maintained. The recombinants were electroporated into BCG to construct the recombinant BCG vaccine and induced by heating. The induced expression detected by SDS-PAGE showed that the content of 65 kD protein expressed in recombinant BCG was 35.69 % in total bacterial protein and 74.09 % in the cell lysate supernatants, suggesting that the recombinant HSP65 gene could express in BCG with high efficiency and the expressed proteins were mainly soluble. Western-blot showed that the secretive recombinant proteins could specifically combine with antibody against M. tuberculosis HSP65, indicating that the recombinant proteins possess the biological activity of HSP65.

Screening Peptide Inhibitors Using Phage Peptide Library with Isocitrate Lyase in Mycobacterium tuberculosis as Target

When devoured by macrophages,Mycobacterium tuberculosis remains persistent in macrophages and gains energy through the glyoxylate bypass to maintain its long-term existence in host cells.Therefore it is possible to stop persistent infections by interdicting the glyoxylate bypass in which the isocitrate lyase（ICL） is the key rate-limiting enzyme and a persistence factor.ICL is the target of anti-TB（TB：tubercular） drugs,which could screen ICL out and effectively inhibit the activity of ICL in Mycobacterium tuberculosis,and because of this,anti-TB drugs can be used to kill persistent Mycobacterium tuberculosis.In this study,the ICL gene of the Mycobacterium tuberculosis H37Rv was cloned successfully and recombinant protein with bioactivity was obtained through the enzyme characteristic appraisal.The specific activity of the recombined ICL is 24μmol·mg-1·min-1.The recombined ICL protein was used as the target,and phages which can specifically combine to ICL were screened in the phage 7 peptide library.According to the results of the ELISA and DNA sequence detection,eventually three 7-peptide chains were synthesized.Then the peptide chains were reacted with ICL,respectively,to detect their inhibitory effects on ICL.The results show that all the three 7-peptide chains possessed varying inhibitory effects on the activity of ICL.This study provided lead compounds for the research and development of new peptide anti-TB drugs.

尘肺结核患者Mycobacterium tuberculosis耐药基因突变研究

研究尘肺结核患者结核分枝杆菌(Mycobacterium tuberculosis,MTB)耐药基因突变与耐药性的关系。在97例尘肺结核患者痰中检出MTB菌株28株,采用PCR-SSCP法检测katG、rpoB和rpsL基因突变,并与采用常规药敏试验(AST)法检测的耐药结果进行对比分析。采用PCR-SSCP法检测katG、rpoB和rpsL基因,突变率分别为42.86%(12/28)、42.86%(12/28)和32.14%(9/28),采用AST法检测INH、RFP和SM,耐药率分别为64.23%(18/28)、60.71%(17/28)和53.57%(15/28),两者之间差异均无显著性(P>0.05)。其中多耐药株20例(71.43%),包括耐三种药物12例(42.86%),耐两种药8例(28.57%),单耐药株6例(21.43%),敏感株2例,耐药率92.86%。PCR-SSCP法检出3个基因联合突变8株,与AST法符合率为66.67%(8/12);2个基因突变5株,符合率为62.50%(5/8);单基因突变2株,符合率为33.33%(2/6)。结果表明:PCR-SSCP技术适用于MTB katG、rpoB和rpsL耐药基因突变的筛选,对指导尘肺结核患者临床用药上具有重要意义。

Study on Mutation and Its Characteristics of Mycobacterium Tuberculosis Multidrug Resistance Genes Based on Whole-genome Sequencing

Objective: The increase in the development of resistance to multiple drugs in mycobacterium tuberculosis(MTB) poses a substantial obstacle to the prevention and management of tuberculosis(TB). A thorough investigation of the genotypes linked to multidrug resistance is crucial for comprehending the mechanisms underlying drug resistance. The objective of this research was to assess the attributes of gene mutations associated with multidrug resistance in clinical isolates of mycobacterium tuberculosis through the utilization of whole-genome sequencing. Methods: A total of 124 strains of drug-resistant mycobacterium tuberculosis were collected, and the genomic DNA of both multidrug-resistant and rifampin-resistant strains were extracted and sequenced. Bioinformatics was used to analyze and compare multidrug resistance-related gene sequences in order to detect the variation of multidrug resistance genes. Results: The results revealed that the resistance spectrum of XDR-TB group was much wider than that of the other three groups, with the RR-TB group having the most limited resistance spectrum.Within the MDR-TB strains, fabG1 exhibited the highest frequency of mutations, while RRS, gyrA, and rpoB were identified as the predominant mutation bases in XDR-TB strains. Additionally, rpoB emerged as the primary mutation base in MDR-TB and RR-TB strains. Notably, the fabG1 mutation was found to be closely associated with PDR-TB. Furthermore, the correlation between the mutation rate of rpoB and multidrug resistance was deemed to be of secondary importance. Conclusion: Various strains of MTB exhibited distinct mechanisms of drug resistance, with the gene mutations of fabG1,RRS, gyrA, and rpoB potentially playing a pivotal role in the development of drug resistance. However, the primary genes responsible for drug resistance mutations varied among different strains of TB.

Effects of Mycobacterium vaccae vaccine in a mouse model of tuberculosis: protective action and differentially expressed genes

Background:Tuberculosis is a leading cause of death worldwide.BCG is an effective vaccine,but not widely used in many parts of the world due to a variety of issues.Mycobacterium vaccae(M.vaccae)is another vaccine used in human subjects to prevent tuberculosis.In the current study,we investigated the potential mechanisms of M.vaccae vaccination by determining differentially expressed genes in mice infected with M.tuberculosis before and after M.vaccae vaccination.Methods:Three days after exposure to M.tuberculosis H37 Rv strain(5×10~5 CFU),adult BALB/c mice randomly received either M.vaccae vaccine(22.5μg)or vehicle via intramuscular injection(n=8).Booster immunization was conducted 14 and 28 days after the primary immunization.Differentially expressed genes were identified by microarray followed by standard bioinformatics analysis.Results:M.vaccae vaccination provided protection against M.tuberculosis infection(most prominent in the lungs).We identified 2,326 upregulated and 2,221 downregulated genes in vaccinated mice.These changes could be mapped to a total of 123 signaling pathways(68 upregulated and 55 downregulated).Further analysis pinpointed to the MyD88-dependent TLR signaling pathway and PI3 K-Akt signaling pathway as most likely to be functional.Conclusions:M.vaccae vaccine provided good protection in mice against M.tuberculosis infection,via a highly complex set of molecular changes.Our findings may provide clue to guide development of more effective vaccine against tuberculosis.

Using a cDNA microarray to study cellular gene expression altered by Mycobacterium tuberculosis

Objective To examine the global effects of Mycobacterium tuberculosis ( M tuberculosis ) infection on macrophages Methods The gene expression profiling of macrophage U937, in response to infection with M tuberculosis H 37 R a, was monitored using a high density cDNA microarray Results M tuberculosis infection caused 463 differentially expressed genes, of which 366 genes are known genes registered in the Gene Bank These genes function in various cellular processes including intracellular signalling, cytoskeletal rearrangement, apoptosis, transcriptional regulation, cell surface receptors, cell mediated immunity as well as a variety of cellular metabolic pathways, and may play key roles in M tuberculosis infection and intracellular survival Conclusions M tuberculosis infection alters the expression of host cell genes, and these genes will provide a foundation for understanding the infection process of M tuberculosis The cDNA microarray is a powerful tool for studying pathogen host cell interaction

Reprogramming Mycobacterium tuberculosis CRISPR System for Gene Editing and Genomewide RNA Interference Screening

Mycobacterium tuberculosis is the causative agent of tuberculosis(TB), which is still the leading cause of mortality from a single infectious disease worldwide. The development of novel anti-TB drugs and vaccines is severely hampered by the complicated and time-consuming genetic manipulation techniques for M. tuberculosis. Here, we harnessed an endogenous type Ⅲ-A CRISPR/Cas10 system of M. tuberculosis for efficient gene editing and RNA interference(RNAi).This simple and easy method only needs to transform a single mini-CRISPR array plasmid, thus avoiding the introduction of exogenous protein and minimizing proteotoxicity. We demonstrated that M. tuberculosis genes can be efficiently and specifically knocked in/out by this system as confirmed by DNA high-throughput sequencing. This system was further applied to single-and multiple-gene RNAi. Moreover, we successfully performed genome-wide RNAi screening to identify M. tuberculosis genes regulating in vitro and intracellular growth. This system can be extensively used for exploring the functional genomics of M. tuberculosis and facilitate the development of novel anti-TB drugs and vaccines.

Exploring Mycobacterium tuberculosis infection-induced alterations in gene expression in macrophage by microarray hybridization

Tuberculosis remains a serious threat to public health. Its causative agent Mycobacte- rium tuberculosis is an intracellular pathogen which survives and replicates within cells of the host immune system, primarily macrophages. Knowledge of the bacteria-macrophage interaction can help to develop novel measures to combat the disease. The global gene expression of macro- phage following invasion by and growth of M. tuberculosis was studied by cDNA microarray. Of the 12800 human genes analyzed, totally 473 (3.7%) macrophage genes were differentially expressed after being infected by M. tuberculosis, among which, only 25 (5.2%, corresponding to less than 0.2% of the 12800 genes) genes were up-regulated, while others (94.8%) were down-regulated against the control. Of the 473 genes, 376 genes are registered in the GenBank, and 97 are novel genes. Expression of 5 up-regulated genes has been induced by more than 3-fold. 25 genes were down-regulated by more than 3-fold. Syndecan binding protein has been down-regu- lated up to 12.5-fold. The data gave an insight into the early gene expression in macrophage ensuing M. tuberculosis infection and a basis for further study.

Rapid Detection of rpoB Gene Mutations in Rif-resistant M.tuberculosis Isolates by Oligonucleotide Microarray

Objective To detect the specific mutations in rpoB gene of Mycobacterium tuberculosis by oligonucleotide microarray. Methods Four wild-type and 8 mutant probes were used to detect rifampin resistant strains. Target DNA of M. tuberculosis was amplified by PCR, hybridized and scanned. Direct sequencing was performed to verify the results of oligonucleotide microarray Results Of the 102 rifampin-resistant strains 98 （96.1%） had mutations in the rpoB genes. Conclusion Oligonucleotide microarray with mutation-specific probes is a reliable and useful tool for the rapid and accurate diagnosis of rifampin resistance in M. tuberculosis isolates.

The bacterial and host factors associated with extrapulmonary dissemination of Mycobacterium tuberculosis

With high morbidity and mortality worldwide, tuberculosis （TB） is still an important public health threat. The majority of human TB cases are caused by Mycobacterium tuberculosis. Although pulmonary TB is the most common presentation, M. tuberculosis can disseminate into other organs and causes extrapulmonary TB （EPTB）. The dissemination of bacteria from the initial site of infection to other organs can lead to fatal diseases, such as miliary and meningeal TB. Thoroughly understanding the mechanisms and pathways of dissemination would develop therapies to prevent the lethal prognosis of EPTB （miliary and meningeal TB） and vaccines to promote the development of adaptive immunity. This review focuses on risk factors of EPTB, bacterial and host genes involved in EPTB, and potential mechanisms of M. tuberculosis extrapulmonary dissemination.

Gene X-pert MTB/RIF检测对肺结核病的诊断价值研究

目的 研究分析肺结核病应用Gene X-pert MTB/RIF检测的诊断价值。方法 选取2022年3—10月在本院就诊的疑似肺结核病患者164例为研究对象,所有患者均给予结核分枝杆菌培养检查、痰涂片抗酸染色法诊断、Gene X-pert MTB/RIF检测,以结核分枝杆菌培养检查结果为金标准,比较痰涂片抗酸染色法诊断与Gene X-pert MTB/RIF检测的阳性率以及诊断准确性、灵敏度、特异度。结果 痰涂片抗酸染色法诊断阳性率为21.34%(35/164),Gene X-pert MTB/RIF检测阳性率为35.98%(59/164),Gene X-pert MTB/RIF检测明显高于痰涂片抗酸染色法诊断(P<0.05)。痰涂片抗酸染色法诊断的准确性为85.37%,灵敏度为59.65%,Gene X-pert MTB/RIF检测的准确性为95.12%,灵敏度为94.74%,Gene X-pert MTB/RIF检测明显高于痰涂片抗酸染色法诊断(P<0.05)。结论 在肺结核病诊断中,Gene X-pert MTB/RIF检测的阳性率更高,同时诊断准确率与灵敏度也更高,为肺结核病的诊断与治疗提供了可靠的指导依据。

广州地区利奈唑胺耐药结核分枝杆菌耐药基因特征分析

目的分析广州地区利奈唑胺(linezolid,LZD)耐药结核分枝杆菌临床分离株耐药基因的突变特征,为控制广州地区耐利奈唑胺结核病提供一定的研究基础和依据。方法选取2012年1月~2022年1月广州市胸科医院住院患者的60株利奈唑胺耐药结核分枝杆菌的临床分离株,分为耐多药组和广泛耐药组。采用微孔板稀释法对60株菌株进行利奈唑胺最低抑菌浓度(MIC)检测,并对60株菌株的利奈唑胺耐药基因23S rRNA、rplC、rplD进行测序,分析其突变特征。结果60株LZD耐药株对LZD的MIC值分布于2~32μg/ml之间。耐多药组的MIC50和MIC90分别为2μg/ml、32μg/ml,广泛耐药组的MIC50和MIC90分别为8μg/ml、32μg/ml。60株菌株中发现有12株23S rRNA基因发生突变,其中有1株发生两个基因位点突变;有11株rplC基因发生突变,其中有1株发生双位点突变;发现2株rplD基因发生突变。23S rRNA、rplC基因突变发生率在不同性别和年龄患者中差异无统计学意义(P>0.05)。结论广州地区利奈唑胺耐药结核分枝杆菌耐药基因23S rRNA、rplC突变发生率明显高于rplD,在不同性别和不同年龄患者中的发生率无显著差异。

Gene Xpert-MTB/RIF检测法在结核病诊断中的意义

目的探讨Gene Xpert-MTB/RIF检测结核分枝杆菌及利福平耐药性在结核病诊断中的作用。方法收集2017年8月至2017年12月在我院住院的202例结核病患者,对其痰标本分别行痰涂片、痰培养、比例法体外药敏试验和Gene Xpert-MTB/RIF法检测,并对Xpert-MTB/RIF检测结果进行分析。结果 Xpert MTB/RIF阳性率及痰培养阳性率明显高于痰涂片,差异有统计学意义;而Xpert MTB/RIF阳性率与痰培养阳性率比较,差异无统计学意义;以痰培养为金标准,Xpert MTB/RIF检测结核分枝杆菌及利福平耐药的敏感度分别为75.9%和66.7%,特异度分别为82.8%和100.0%。Xpert MTB/RIF及传统比例法检测利福平耐药一致性好。结论 Xpert检测与传统方法相比快捷,并具有更高灵敏度及特异度,可为临床诊断、选择耐药结核的治疗方案提供参考。

Gene MTB/RIF、TB-IGRA检测与传统组织病理学检查在脊柱结核诊断中的应用价值

目的:评估传统组织病理学(简称“病理”)检查与Gene MTB/RIF(简称“Xpert”)、γ干扰素释放试验(TB-IGRA)在脊柱结核诊断中的应用价值。方法:回顾性分析2017年11月~2020年6月在我院经临床诊断为“脊柱结核”的131例患者资料,男79例,女52例;年龄18~90岁(50±18.0岁);颈椎7例,胸椎39例,胸腰段11例,腰椎57例,腰骶椎12例,骶椎5例。所有患者术前进行TB-IGRA检测,然后通过穿刺或手术获得病灶组织,分别进行病理检查与Xpert检测。病理检查结果分四类:Ⅰ类为确诊结核、Ⅱ类为倾向于结核,Ⅲ类为疑诊结核,Ⅳ类为未确诊结核。Ⅰ类、Ⅱ类病理结果支持脊柱结核诊断,Ⅲ类、Ⅳ类不支持脊柱结核诊断。计算三者的阳性率与阴性率。以病理检查为参照标准,计算Xpert、TB-IGRA的敏感度、特异度,三者单独检测的阳性率、联合检测的阳性率分别进行比较,计算Kappa值评估两者的一致性,并绘制Xpert、TB-IGRA检测的受试者工作特征(receiver operating characteristic,ROC)曲线并计算曲线下面积(area under curve,AUC),评估Xpert、TB-IGRA检测的价值。结果:所有患者中,病理确诊为结核85例(64.9%,95%CI为56.6%~73.2%),病理未确诊结核46例(35.1%,95%CI为26.8%~43.4%);Xpert检测阳性79例(60.3%,95%CI为51.0%~68.1%),阴性52例(39.7%,95%CI为31.9%~49%),发现RNA聚合酶β亚基的编码基因(rpoB)突变6例(4.6%,95%CI为0.95%~8.20%);TB-IGRA检测阳性99例(75.6%,95%CI为68.1%.83.0%),阴性32例(24.4%,95%CI为17.0%~31.9%)。病理检查与Xpert检测联合诊断脊柱结核89例(67.9%,95%CI为59.02%~75.32%),未确诊结核42例(32.1%,95%CI为24.68%~40.98%)。病理检查与TB-IGRA检测联合诊断脊柱结核107例(81.7%,95%CI为74.97%~88.39%),未确诊结核24例(18.3%,95%CI为11.61%~25.03%)。Xpert检测敏感度为87.1%(74/85),特异度为91.3%(42/46);TB-IGR检测敏感度为90.6%(77/85),特异度为52.2%(24/46)。病理检查与Xpert检测联合诊断脊柱结核的阳性率为67.9%(89/131);病理检查与TB-IGRA检测联合诊断脊柱结核的阳性率为81.7%(107/131);三者联合诊断脊柱结核的阳性率为82.4%(108/131)。以临床诊断结果作为参照,TB-IGRA联合病理检查阳性率高于病理检查(χ^(2)=9.435,P=0.002),三者联合检测阳性率高于病理检查(χ^(2)=9.855,P=0.002),也高于Xpert与病理检查联合(χ^(2)=16.681,P<0.001)。病理检查与Xpert两种检测确诊脊柱结核的Kappa值为0.76(95%CI为0.631~0.873),一致性好;病理检查与TB-IGRA两种检测确诊脊柱结核的Kappa值为0.46(95%CI为0.295~0.616),一致性较好。Xpert检测诊断脊柱结核的AUC为0.892;TB-IGRA检测诊断脊柱结核的AUC为0.751。结论:TB-IGRA检测敏感性较高,Xpert检测特异性较高,并且能发现利福平耐药突变;二者联合病理学检查对诊断脊柱结核具有较高的应用价值。

DNA-微阵列芯片检测技术与Gene-Xpert MTB/RIF技术在新疆维吾尔自治区结核分枝杆菌耐药性诊断中的应用

目的探索DNA-微阵列芯片检测技术(以下简称基因芯片技术)和利福平耐药实时荧光定量核酸扩增检测技术(Gene-Xpert MTB/RIF技术)检测新疆维吾尔自治区结核分枝杆菌中利福平和异烟肼耐药性的应用价值。方法收集2015年新疆维吾尔自治区结核病耐药监测菌株115株,对所有的菌株进行比例法药敏试验、基因芯片技术、Gene-Xpert MTB/RIF技术检测。以比例法药敏试验为“金标准”,分别计算基因芯片技术与Gene-Xpert MTB/RIF技术对结核分枝杆菌耐药性诊断的效能并进行评价。结果比例法药敏试验结果显示,利福平耐药患者40例(34.78%),异烟肼耐药患者48例(41.74%)。以比例法药敏试验结果为“金标准”,基因芯片技术检测结核分枝杆菌对利福平和异烟肼耐药耐药性诊断的灵敏度、特异度分别为80.00%、89.33%和77.08%、97.01%,Gene-Xpert MTB/RIF技术检测结核分枝杆菌对利福平耐药性诊断的灵敏度、特异度分别为92.50%、94.67%。基因芯片技术和Gene-Xpert MTB/RIF技术对利福平耐药患者的诊断效能差异无统计学意义(P>0.05)。结论对于新疆维吾尔自治区结核病耐药监测菌株,基因芯片技术和Gene-Xpert MTB/RIF技术对耐药结核病的诊断均具有较高的灵敏度和特异度。Gene-Xpert MTB/RIF技术操作方法简单,检测时间短;基因芯片技术耐药检测范围广,耗时少,为新疆维吾尔自治区广泛开展耐药结核病快速检测提供了有力的技术支持。

纳米孔靶向测序在分枝杆菌鉴定和耐药基因检测中的研究进展

2023年全球结核病报告显示,结核病的流行病学形势依然严峻。现有的检测方法在分枝杆菌菌种鉴定和耐药基因检测方面存在不足之处,临床亟需开发更加便捷、高效、准确的检测技术,以满足结核病早期诊断和指导治疗的需求。纳米孔靶向测序具有便捷高效、长读长测序等优势,给结核病和耐药结核病的诊疗带来了新的契机。笔者就纳米孔靶向测序这一新技术的原理与特点,以及在分枝杆菌鉴定和耐药基因检测方面的应用进行综述,旨在为临床工作者探索新技术提供参考。