聚合酶链反应技术诊断泌尿系结核

采用聚合链反应（ＰＣＲ）技术快速检测尿液标本中结核相菌ＤＮＡ，显示敏感性为９２％特异性为８８％，阳性预测７９％，阴性预测９６％，可靠性为８９％，认为ＰＣＲ是临床快速诊断泌尿系结核的有效方法之一，优于现行病原学检查法，适合临床推广应用。     

聚合酶链反应对关节结核病诊断与鉴别诊断的临床价值

为探讨聚合酶链反应对关节结核病诊断与鉴别诊断的临床价值，对３０例关节结核和２０例非关节结核标本分别采用ＰＣＲ，抗酸染色镜检与分离增减法检测结核杆菌。结果３０例关节结核标本中阳性检率分别为ＰＣＲ地８７％，镜检法０，培养法１３％。     

聚合酶链反应检测结核杆菌DNA对关节结核的诊断价值

聚合酶链反应（以下简称ＰＣＲ）是近几年发展起来的基因诊断技术。我们自１９９４年开始对２６例结核性关节液进行涂片染色分析，结核杆菌培养以及ＰＣＲ检测，并与３５例非结核性关节液的ＰＣＲ检测相对照，就其结果分析如下。临床资料结核性关节液２６例中男１６例，女１０例；年龄１５～６８岁；关节部位：髋关节６例，膝关节１３例，踝关节２例，肘关节３例，腕关节２例，以膝关节为最多。均依据病史、临床表现、Ｘ线检查及实验室检查，并经手术、病理确诊为骨关节结核。非结核性关节液３５例中男２１例，女１４例；年龄１１～６５岁。…     

DNA体外扩增技术诊断关节结核的临床研究

目的：为探讨ＤＮＡ体外扩增（ｐｏｌｙｍｅｒａｓｅｃｈａｉｎｒｅａｃｔｉｏｎ，ＰＣＲ）技术对关节结核诊断的临床价值。材料与方法：对３０例关节结核和２０例非关节结核标本分别应用ＰＣＲ技术、抗酸染色镜检及分离培养法检测结核杆菌。结果：３０例关节结核中三种方法的阳性检出率分别为：ＰＣＲ法８７％，镜检法０，培养法１３％，ＰＣＲ法与镜检及培养法对于结核杆菌的阳性检出率比较，差异具有显著性意义（Ｐ值＜０．００５），ＰＣＲ法明显优于镜检及培养法。２０例非关节结核标本镜检及培养法均阴性，ＰＣＲ法阳性率１０％。盲法结核杆菌和对照菌ＰＣＲ检测结果表明，ＰＣＲ的特异性为１００％。ＰＣＲ扩增整个过程自动化控制，可在数小时内完成。结论：此表明ＰＣＲ技术检测关节结核标本结核杆菌具有快速、简便、特异与敏感等优点，对关节结核的早期快速诊断与鉴别诊断具有重要价值。     

临床诊断骨与关节结核患者病原菌的培养鉴定及耐药分析

【摘要】 目的：分析临床诊断骨与关节结核患者病原菌的种类及耐药情况,为骨与关节结核的诊断和治疗提供参考。方法：收集2008年12月~2009年9月在新疆医科大学第一附属医院临床诊断骨与关节结核而接受手术治疗患者的病灶标本69例,采用BACTEC MGIT 960系统进行分枝杆菌培养、菌种鉴定和药物敏感试验,分析分枝杆菌培养阳性率、菌种分布及耐药情况。结果：在培养4周内24例（34.78%）分枝杆菌培养阳性,延长至8周时共有40例（57.97%）培养阳性,两者比较有统计学差异（P＜0.05）;其中结核分枝杆     

聚合酶链反应检测结核杆菌DNA对骨结核病诊断的临床价值

采用聚合酶链反应（ＰＣＲ），分别对２０例骨结核患者标本用两种方法检查的阳性率分别为：培养法２０％，ＰＣＲ６５％，经统计学处理二种方法有显著性差别，ＰＣＲ阳性率显著高于培养法，而且２天即可报告，显著缩短了报告时间。３０例非骨结核中患者标本两种方法均为阴性。盲法结核杆菌和对照菌ＰＣＲ结果表明具有较高的特异性。只要标本中有结核杆菌ＤＮＡ即可扩增其特异的ＤＮＡ片断，即使经福尔马林固定２－４周的酝酿也可检测     

突变体富集聚合酶链反应检测胰腺癌K-ras基因突变的研究

胰腺癌是一种恶性程度极高，早期难以诊断，预后极差的恶性肿瘤，临床确诊者大多属于晚期癌，5年生存率总体低于5％。我们采用突变体富集聚合酶链反应（PCR）检测胰腺癌细胞系K—ras基因点突变，以期为早期诊断胰腺癌提供一种简便、灵敏而有效的检验方法。     

Clinical value of polymerase chain reaction in the diagnosis of joint tuberculosis by detecting the DNA of Mycobacterium tuberculosis

Objective: To assess the clinical value of polymerase chain reaction (PCR) in the diagnosis and differential diagnosis of joint tuberculosis (TB). Methods: PCR was used blindly to detect the DNA of Mycobacterium tuberculosis (M.TB) in five specimens of M.TB, 5 of BCG, and 10 of other bacteria. Then, M. TB in 98 samples from patients with joint TB and 100 samples from patients with non‐tubercular joint disorders were detected by PCR, acid‐fast staining and culture,. The sensitivity, specificity, accuracy, positive predictive value, and negative predictive value of PCR were calculated. The χ2 test was used for statistical analysis of the frequency of various factors. At the same time, some problems with PCR were also systematically analyzed. Results: (1) In the “standard samples”, both M. TB and BCG showed positive while other bacteria were negative. (2) In 98 cases from patients with joint TB, 81 were positive by PCR, 6 by acid‐fast staining, and 17 by culture. In 100 cases from patients with non‐tuberculous joint disorders, 9 were positive by PCR, and none by either acid‐fast staining or culture. Sensitivity, specificity, accuracy, positive and negative predictive value of PCR were 82.65% (81/98), 91.00% (91/100), 86.87% (172/198), 90.00% (81/90) and 84.26% (91/108), respectively. (3) The positive rates for PCR, acid‐fast staining and culture in detection of M. TB were 82.65% (81/98), 6.12% (6/98), and 17.34% (17/98), respectively. There were statistically significant differences between the three methods (P < 0.001). (4) The process of PCR is automatic, and can be completed within 3 to 6 hours, whereas 4 to 8 weeks are required for the conventional culture of M. TB. Conclusion: PCR is a sensitive, specific, rapid, simple and minimally invasive method for detection of M. TB in samples from joint TB, and can play an important role in early and rapid diagnosis and differential diagnosis of joint TB. But it also has some limitations, such as false positivity and false negativity.     

PCR检测尿结核杆菌的临床应用价值

评价聚合酶链反应（ＰＣＲ）诊断泌尿系结核的临床应用价值，采用ＰＣＲ检测经病理证实为泌尿系结核９例，并报告１７例尿ＰＣＲ阳性的可疑肾结核抗痨治疗前后的临床情况，结果：９例泌尿系结核者，ＰＣＲ检测阳性７例；１７例尿ＰＣＲ阳性的可凝肾结核者经抗痨治疗３个月后，有１４例症状完全消失，尿常规转为正常，认为ＰＣＲ是临床快速诊断泌尿系结核的有效方法。     

多聚酶链反应检测尿中结核杆菌的临床意义

应用多聚酶链反应（ＰＣＲ）对８６例患者（１０例经病理诊断为肾结核，６９例可疑肾结核，７例单纯附睾结核）和３０例健康对照者进行连续２日晨尿结核菌检测。１０例肾结核患者检出均阳性；可疑肾结核者第一次检出９例，第二次为６例；７例附睾结核者两次无一阳性；对照组有１例二次检查均为阳性。认为ＰＣＲ对尿中结核杆菌检出率高、准确、快速，值得在临床上推广应用。     

骨关节结核病灶中耐多药结核分枝杆菌对疗效的影响

目的回顾性探索骨关节结核病灶中耐多药结核分枝杆菌对骨关节结核病疗效的影响及其对策。方法1993至1999年,确诊为复治骨关节结核病250例,男143例,女107例;年龄2～72岁,平均25.44岁;病变在脊椎、髋、膝和其他小关节。穿刺或外科手术获得病灶中脓液、干酪样物或肉芽组织等为标本,以BACTECTB460检测仪进行结核菌培养和药敏试验,NAP试剂进行初级鉴定,NTM分离培养物进一步鉴定分枝杆菌菌种。结果250例标本中,分离培养阳性培养物48例。结核分枝杆菌46例,阳性率为18.4%。其中同时耐异烟肼(INH)、利福平(RFP)、链霉素(SM)和乙胺丁醇(EMB)中三种药的结核分枝杆菌4例,同时耐异烟肼、利福平、链霉素和乙胺丁醇11例,共15例(6%)。在15例中有8例此次手术后切口破溃形成窦道,依据药敏试验结果改变化疗方案治疗,并经前后2或3次再手术,7例治愈,1例因窦道继发感染而死亡。另2例为非结核分枝杆菌(NTM),1例鉴定为鸟胞内分枝杆菌复合群,另1例鉴定未有结果,但均为耐多药(INH、RFP、SM、EMB、KM、CS)的NTM,治疗效果差。结论骨关节结核病灶中发现耐多药结核分枝杆菌(6%)和非结核分枝杆菌的病例,疗效极差,此类病例的化疗方案应个体化,可能需要多次手术才能治愈。     

PCR诊断早期肾结核的探讨

探讨聚合酶链反应（ＰＣＲ）诊断早期肾结核的实用价值，应用ＰＣＲ技术快速检测尿中结核杆菌ＤＮＡ诊断不明原因的无症状性镜下血尿者２３例，结果９例阳性，经抗结核治疗，９例镜下血尿全部消失，认为ＰＣＲ技术诊断早期肾结核具有速度快、特异性高，灵敏度大的优点，可列为疑诊早期肾结核检测常规。     

关节镜技术在青少年膝关节结核诊治中的初步应用

目的 总结关节镜技术在青少年膝关节结核诊断和治疗中的经验.方法 2002年6月至2006年12月,对36例诊断为结核,5例不具备结核诊断标准,按滑膜炎待查的患儿采用膝关节镜下清理及抗结核药物治疗,男13例,女28例;年龄7~16岁,平均12.5岁;均有不同程度的膝关节肿胀、轻度疼痛及功能障碍;病史3~14个月,平均4.8个月.随访时利用Lysholm进行问卷调查,同时对其临床资料进行回顾性分析.结果 41例患儿,术后病理共确诊37例,另4例报告为滑膜组织慢性炎症或坏死组织.随访6~58个月,平均43.5个月,所有患膝经关节镜和术后药物治疗均无复发.Lysholm评分75~98分,平均91.4分.6例患儿残留不同程度膝关节活动受限,活动范围70°~110°,其中2例全关节结核伸直滞缺分别为10°和25°.结论 青少年膝关节结核的诊治较为特殊,在病程早期采用膝关节镜下清理、药物治疗结合术后早期康复锻炼的方案具有创伤小、病灶清除彻底等优点,并可最大限度保留膝关节功能,避免外科介入过晚而导致的膝关节融合,是一种较为理想的治疗方法.     

聚合酶链技术在慢性前列腺炎诊治中的应用

目的：探索慢性前列腺炎的病因，提高其诊治水平。方法：应用聚合酶链技术(PCR)对门诊疑诊为慢性前列腺炎患者的前列腺液进行淋球菌(NG)、沙眼衣原体(CT)及解脲支原体(UU)检测。结果：检出单纯淋球菌感染33例，阳性率9．09％；沙眼衣原体感染32例，阳性率5．15％；解脲支原体感染250例，阳性率23．00％，沙眼衣原体与解脲支原体合并感染13例；淋球菌合并解脲支原体感染5例；淋球菌合并沙眼衣原体感染4例；并应用PCR监测治疗效果。结论：PCR检测慢性前列腺炎病原体快速、敏感性高、特异性强，是一种较为理想、值得推广的临床应用技术。     

基于85B mRNA的逆转录-实时定量聚合酶链反应对肺结核分枝杆菌感染的诊断价值

目的探讨痰液样本中检测结核分枝杆菌（MTB）85B mRNA的逆转录-实时定量聚合酶链反应（RT-qPCR）对MTB感染的诊断价值。 方法分别纳入2016年1月至2016年12月陕西省结核病防治院经痰培养法（BACTEC MGIT 960系统）确诊为MTB阳性患者60例为试验组,60例无MTB感染的健康人为对照组。提取痰液样本核酸,分别采用TaqMan法检测MTB基因中编码16S rRNA的DNA序列和RT-qPCR法检测MTB的85B mRNA,比较两种方法的诊断效能（敏感性、特异性、阳性预测值、阴性预测值和准确性）。 结果TaqMan法诊断MTB感染的敏感性、特异性、阳性预测值、阴性预测值和准确性分别为93.3%、83.3%、84.9%、92.6%和88.3%;RT-qPCR法分别为98.3%、95.0%、95.2%、98.3%和96.7%。通过二元Logistics回归分析证实基于85B mRNA的RT-qPCR法对MTB感染鉴别能力更强（OR = 19.924,95%CI：5.364～73.012,P < 0.001）。 结论基于结核分枝杆菌85B mRNA的RT-qPCR方法能够快速、有效诊断MTB感染。     

Polymerase chain reaction in the diagnosis of urinary tract tuberculosis

The polymerase chain reaction (PCR) is a technique that can be used to amplify a specific DNA genomic sequence, whereby the presence of an extremely small number of bacteria can be detected. The high sensitivity of PCR is particularly useful in paucibacillary situations such as non-pulmonary tuberculosis (TB). The aims of the present study were to establish a PCR assay for the rapid detection of Mycobacterium tuberculosis (MTb) in urine, to compare the sensitivity of PCR with routine culture technique (Bactec) and to determine the optimal type of urine specimen for PCR detection of MTb. In the first phase of the study, a total of 92 urine specimens were collected from 83 patients with suspected urinary tract TB. Two urine specimens in 2 patients were positive for TB by both PCR and Bactec, while 90 specimens from 81 patients were negative by both methods. Inhibition of PCR was present in nine urine specimens (10%). In the second phase of the study, a further seven patients were selected for intensive investigation to determine the optimal urine sampling for PCR detection of MTb. The conclusions of the study are that PCR can provide much faster confirmation of urinary TB (within 24–48 h) than Bactec urine culture (which may take several weeks). About 10% of urine specimens could not be evaluated by PCR due to the presence of inhibitory substances of unknown nature. MTb organisms were found to be excreted intermittently in the urine of infected patients, and single specimens were more likely to be false negative than a 24-h sample. The best method appeared to be the concentration of a large volume of urine, for instance 11 concentrated to 2 ml.     

Comparison of polymerase chain reaction and bacteria culture in peritoneal dialysis associated peritonitis

Objective To evaluate clinical application value of polymerase chain reaction (PCR) detection for bacteria in peritoneal dialysis associated peritonitis (PDAP). Methods Peritoneal dialysis fluid specimens were collected from January 2014 to December 2014 in The First Affiliated Hospital of Anhui Medical University. Conventional bacterial culture and PCR detection were used respectively. According to the bacterial 16S rRNA gene, universal primers were devised and designed, based on reference, the specific primers of 17 kinds of experimental bacteria. Real-time fluorescent PCR (Real-time PCR, qPCR) amplification was implemented. The establishment of standard strain DNA extract was used as positive control; sterile double distilled water was used as negative control. Results (1) The traditional bacterial culture results showed that positive proportion was 26/40 in specimen of 40 cases, gram-positive strains accounting for 18/26. Main species were epidermis staphylococcus (5/26), hemolysis staphylococcus (4/26), escherichia coli (4/26), and streptococcus viridans (3/26). (2) The PCR detection results showed that total positive rate was 33/40 in 40 patients specimens, among which 2 cases of positive samples ended up with no specific strains being detected; the main bacteria strains in PCR were not different from ordinary culture results. (3) With bacterial culture as the gold standard, the detection sensitivity of PCR technology for PDAP pathogenic bacteria was 96.15% and specificity was 42.86%; the detection positive rate was significantly higher than ordinary culture method. (4) PCR technology for detecting pathogenic bacteria could produce results within 4-6 hours, while reported positive results in the traditional bacterial culture would take (77.88±15.53) hours, which was significantly longer than PCR. Conclusion Compared with traditional bacteria culture method, PCR method is more sensitive, simple, and quick. Bacteria detection using PCR technique is of clinic applied value in PDRP.