本文選題：結(jié)核分枝桿菌　切入點(diǎn)：FQ-PCR　出處：《新鄉(xiāng)醫(yī)學(xué)院》2013年碩士論文

【摘要】：背景 結(jié)核病(tuberclosis)是嚴(yán)重危害人類健康的慢性傳染病,近年來疫情的回升嚴(yán)重威脅著世界各國人民的生命財產(chǎn)安全,已成為全球關(guān)注的重大公共衛(wèi)生問題和社會問題。目前,我國結(jié)核病疫情也呈回升趨勢,并且重癥患者和難治性患者越來越多,呈現(xiàn)出高感染率、高患病率、高死亡率和高耐藥率的特點(diǎn)。因此尋找靈敏、特異、高效、實用的結(jié)核病診斷方法是實驗診斷領(lǐng)域研究的熱點(diǎn),也是結(jié)核病早期診斷、徹底治療、有效預(yù)防和控制蔓延的關(guān)鍵。 結(jié)核病是由結(jié)核分枝桿菌感染引起的,實驗室分離和檢測病原體及其代謝產(chǎn)物是結(jié)核病診斷的主要依據(jù)。常用的結(jié)核桿菌檢測方法雖然各有優(yōu)點(diǎn),但都有一定的局限性,如容易受其它抗酸性分枝桿菌的干擾,檢測靈敏度差、陽性率低、容易造成漏檢。特別是培養(yǎng)法作為目前公認(rèn)的診斷結(jié)核分枝桿菌的金標(biāo)準(zhǔn),仍然存在敏感度低、所需時間過長、對疾病的早期診斷和治療意義不大等不足。近年來,熒光定量PCR(FQ-PCR)結(jié)核桿菌核酸分子(TB-DNA)檢測技術(shù)的建立為結(jié)核病的早期診治和防控帶來了希望。 關(guān)于FQ-PCR在結(jié)核病診斷中的應(yīng)用研究,目前主要集中在對確診病人以上幾種實驗方法的對比方面,在對初次發(fā)病的患者的早期診斷及鑒別診斷方面研究較少,對于不同類型結(jié)核病的相應(yīng)標(biāo)本檢測的有效性方面研究也不多,特別是缺乏大樣本的臨床應(yīng)用評價。鑒于此,我們開展了本實驗。 目的 完善FQ-PCR法對痰液、胸水、腦脊液和尿液等不同類型標(biāo)本TB-DNA的檢測方法,評價其對肺結(jié)核、結(jié)核性胸膜炎、結(jié)核性腦膜炎和泌尿系結(jié)核的早期診斷價值。 方法 隨機(jī)選擇初次就診的可疑結(jié)核病患者為實驗對象,分為肺結(jié)核組、結(jié)核性胸膜炎組、結(jié)核性腦膜炎組和泌尿系結(jié)核組,分別收集痰液、胸水、腦脊液和尿液標(biāo)本,經(jīng)規(guī)范處理后,用FQ-PCR試劑盒進(jìn)行TB-DNA檢測,得到Ct值(cycle threshold),以Ct30并曲線為標(biāo)準(zhǔn)S形曲線的實驗結(jié)果判為陽性。同時進(jìn)行抗酸染色、細(xì)菌培養(yǎng)等實驗室檢查,結(jié)合臨床診療過程和隨訪,確定所選病例的進(jìn)展。按照各類結(jié)核病診斷標(biāo)準(zhǔn)對受試者做出最后診斷。將實驗結(jié)果與最終診斷進(jìn)行比對,做出FQ-PCR對各類結(jié)核病的早期診斷臨床性能評價。 結(jié)果 肺結(jié)核組得到有效資料123例,痰液標(biāo)本FQ-PCR擴(kuò)增TB-DNA陽性66例,陰性57例；金標(biāo)準(zhǔn)判定為肺結(jié)核的93例,不是肺結(jié)核的30例。其中真陽性數(shù)為64,假陽性數(shù)為2,真陰性數(shù)為28,假陰性數(shù)為29。由此計算出該方法的臨床靈敏度尚可,為68.8%,臨床特異度較好,為93.3%,總正確度尚可,為74.8%。陽性預(yù)測值較好,達(dá)96.9%,而陰性預(yù)測值較差,僅49.1%。結(jié)核性胸膜炎組胸水標(biāo)本FQ-PCR擴(kuò)增TB-DNA,其臨床靈敏度、臨床特異度、陽性預(yù)測值和陰性預(yù)測值分別為70.4%(38/54)、96.8%(31/32)、97.4%(38/39)、65.9%(31/47)。結(jié)核性腦膜炎組腦脊液標(biāo)本FQ-PCR擴(kuò)增TB-DNA,其臨床靈敏度、臨床特異度、陽性預(yù)測值和陰性預(yù)測值分別為46.9%(31/66)、92.3%(12/13)、96.9%(31/32)、25.5%(12/47)。泌尿系結(jié)核組受檢的78例疑似病人尿液標(biāo)本,其臨床靈敏度、臨床特異度、陽性預(yù)測值和陰性預(yù)測值分別為38.4%(20/52)、84.6%(22/26)、83.3%(20/24)、40.7%(22/54)。 結(jié)論 FQ-PCR法檢測結(jié)核分枝桿菌對結(jié)核病的早期診斷具有較高臨床價值。痰液和胸水對結(jié)核病的診斷特異性好,陽性預(yù)測值高,臨床靈敏度尚可,但陰性預(yù)測值較低,容易出現(xiàn)漏診。腦脊液和尿液標(biāo)本對相應(yīng)結(jié)核病的診斷有較好的排除性能,但確診性能不夠好。
[Abstract]:background
Tuberculosis (Tuberclosis) is a chronic infectious disease which seriously endangers human health. In recent years, the epidemic rise is a serious threat to the peoples of the world's life and property safety, has become a major public health problem and social problem of the world. At present, the tuberculosis epidemic situation of China also showed increasing trend, and in patients with severe and refractory patients more and more, showing a high infection rate, high morbidity, high mortality and high resistance rate characteristics. So looking for sensitive, specific, efficient and practical method for diagnosis of tuberculosis is a hot research field of experimental diagnosis, it is early diagnosis and treatment of tuberculosis completely, effectively prevent and control the spread of the key.
Tuberculosis is caused by Mycobacterium tuberculosis infection, laboratory of separation and detection of pathogens and its metabolites is the main basis for the diagnosis of tuberculosis. Mycobacterium tuberculosis detection methods commonly used although each have advantages, but there are some limitations, such as vulnerable to other acid resistant Mycobacterium interference, the detection sensitivity is poor, low positive rate, easy to the cause of the undetected. Especially the culture as the gold standard for the diagnosis of Mycobacterium tuberculosis recognized, there are still low sensitivity, it takes long time to disease, early diagnosis and treatment of significant deficiencies. In recent years, fluorescence quantitative PCR (FQ-PCR) of Mycobacterium tuberculosis DNA (TB-DNA) technique for the early detection of the diagnosis and prevention of tuberculosis has brought hope.
Study on Application of FQ-PCR in the diagnosis of tuberculosis, mainly focuses on the comparison of patients above several experimental methods, for the first time in the early onset of the patient's diagnosis and differential diagnosis of small, for the study of the corresponding samples of different types of tuberculosis of the effectiveness is not much, especially the lack of clinical evaluation large sample. In view of this, we carry out this experiment.
objective
To improve the FQ-PCR method of sputum, pleural effusion, cerebrospinal fluid detection method and urine samples of different types of TB-DNA, the evaluation of pulmonary tuberculosis, tuberculous pleurisy, early diagnosis of tuberculous meningitis and urinary tuberculosis.
Method
Random selection of initial treatment of suspected tuberculosis patients as the experimental object, divided into pulmonary tuberculosis group, tuberculous pleurisy group, tuberculous meningitis group and urinary tuberculosis group were collected sputum, pleural effusion, cerebrospinal fluid and urine samples after standard treatment, using the FQ-PCR kit for detection of TB-DNA, Ct value (cycle threshold), with Ct30 and standard curve S curve. The experimental results were considered positive. Acid fast staining at the same time, bacterial culture and laboratory examination, combined with clinical diagnosis and follow-up process, determine the progress of selected cases. According to the diagnostic criteria of all kinds of nodes make the final diagnosis of the disease subjects to compare the experimental results. With the final diagnosis, make FQ-PCR of tuberculosis early diagnosis clinical performance evaluation.
Result
Pulmonary tuberculosis group effective 123 cases sputum samples were amplified by FQ-PCR TB-DNA was positive in 66 cases, 57 cases were negative; the gold standard for determination of 93 cases of pulmonary tuberculosis, 30 cases of pulmonary tuberculosis is not. The true positive rate was 64, false positive rate was 2, true negative rate was 28, the false negative number is 29. calculate the clinical sensitivity is acceptable, the method was 68.8%, the clinical specificity was 93.3%, the total is good, the accuracy is better, up to 96.9%, the positive predictive value of 74.8%., while the negative predictive value is poor, only 49.1%. group of tuberculous pleurisy pleural effusion specimens of FQ-PCR amplification of TB-DNA, the clinical sensitivity, clinical specificity, positive predictive value and negative predictive value were 70.4% (38/54), 96.8% (31/32), 97.4% (38/39), 65.9% (31/47). Tuberculous meningitis cerebrospinal fluid specimens was amplified by FQ-PCR TB-DNA, the clinical clinical sensitivity, specificity, positive predictive value and negative predictive value were 46.9% (31/66), 92.3% (12/13 ), 96.9% (31/32), 25.5% (12/47). Urinary tuberculosis group examined 78 patients with suspected urine specimens, the clinical clinical sensitivity, specificity, positive predictive value and negative predictive value were 38.4% (20/52), 84.6% (22/26), 83.3% (20/24), 40.7% (22/54).
conclusion
Has high clinical value in early diagnosis of tuberculosis detection of Mycobacterium tuberculosis by FQ-PCR. Sputum and pleural effusion diagnosis specificity of tuberculosis, the positive predictive value of high sensitivity, clinical acceptable, but the negative predictive value is low, prone to misdiagnosis. Cerebrospinal fluid and urine specimens for diagnosis of tuberculosis has overcome the corresponding performance. But the diagnosis performance is not good enough.

【學(xué)位授予單位】：新鄉(xiāng)醫(yī)學(xué)院
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2013
【分類號】：R52

【共引文獻(xiàn)】

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本文編號：1717159