本文選題：恒溫?cái)U(kuò)增微控流芯片核酸分析技術(shù) + 人工關(guān)節(jié)置換�。� 參考：《昆明醫(yī)科大學(xué)》2017年碩士論文

【摘要】：人工關(guān)節(jié)置換(Artificial joint replacement,AJR)術(shù)后感染是人工關(guān)節(jié)置換術(shù)后最嚴(yán)重的并發(fā)癥之一,是廣大關(guān)節(jié)外科醫(yī)生不可回避的問題,其診斷的關(guān)鍵是找到致病菌,一旦明確病原菌,便可確定診斷,就可以開始相應(yīng)的治療。目前臨床上主要用于檢測病原菌的方法是細(xì)菌分離培養(yǎng),檢測耗時(shí)長,眾多因素容易造成假陰性結(jié)果的缺點(diǎn)易導(dǎo)致延誤最佳治療時(shí)機(jī)。恒溫?cái)U(kuò)增微控流芯片核酸分析技術(shù)是分子生物學(xué)技術(shù)迅猛發(fā)展背景下應(yīng)運(yùn)而生的新型病原菌檢測技術(shù),具有檢測快速、靈敏度高、準(zhǔn)確性高、簡便高效等優(yōu)點(diǎn)。擯棄了傳統(tǒng)聚合酶鏈反應(yīng)(polymerase chain reaction,PCR)技術(shù)操作繁瑣、溫度要求嚴(yán)格、標(biāo)本易污染的缺點(diǎn)。從標(biāo)本處理開始到檢出結(jié)果,全程只需要2小時(shí)�？梢酝瑫r(shí)檢測多種病原菌。恒溫?cái)U(kuò)增微控流芯片核酸分析技術(shù)目前主要被應(yīng)用于呼吸道病原菌的檢測,在全國5所大型三甲醫(yī)院進(jìn)行的2518例標(biāo)本檢測中,準(zhǔn)確率高達(dá)94.5%。由于此項(xiàng)技術(shù)是近兩年的新興技術(shù),目前尚未應(yīng)用于AJR術(shù)后感染病原菌的檢測,國內(nèi)外也無相關(guān)文獻(xiàn)報(bào)道。本課題通過研究恒溫?cái)U(kuò)增微控流芯片核酸分析技術(shù)在AJR術(shù)后感染病原菌檢測中的效果,探討此項(xiàng)技術(shù)的臨床實(shí)用性及推廣價(jià)值。毋庸置疑,隨著科學(xué)技術(shù)的發(fā)展以及臨床與科研的緊密結(jié)合,恒溫?cái)U(kuò)增技術(shù)在病原菌的診斷中必定占有一席之地。[目的]通過分析恒溫?cái)U(kuò)增微控流芯片核酸分析技術(shù)(Nucleic Acid Analysy Technique of Isothermal Amplification Microfluidic Chip,NAATIAMC)快速檢測AJR術(shù)后感染病原菌的效果,探討其臨床實(shí)用性及推廣價(jià)值。[方法]1.選取2016年1月至2017年2月昆明醫(yī)科大學(xué)第一附屬醫(yī)院骨科及呈貢分院骨科收治的疑似人工關(guān)節(jié)置換術(shù)后感染的28例患者為研究對象,其中男性12例,女性16例,年齡64.5±13.1歲(37～83歲);人工膝關(guān)節(jié)置換術(shù)后8例,人工髖關(guān)節(jié)置換術(shù)后20例。2.將采集到的破潰創(chuàng)口分泌物或關(guān)節(jié)穿刺液、術(shù)中假體周圍組織碾磨液標(biāo)本分為兩組,各取5ml。同時(shí)采用恒溫?cái)U(kuò)增微控流芯片核酸分析技術(shù)(實(shí)驗(yàn)組)和細(xì)菌分離培養(yǎng)法(對照組)進(jìn)行病原菌檢測/培養(yǎng)。3.記錄實(shí)驗(yàn)組和對照組檢測/培養(yǎng)出病原菌的種類及耗時(shí),列出病原菌檢測/培養(yǎng)結(jié)果四格表。通過一致性的假設(shè)檢驗(yàn)比較兩組數(shù)據(jù)結(jié)果的一致性;采用χ2檢驗(yàn),比較兩組陽性率,p0.05,表示差異有統(tǒng)計(jì)學(xué)意義。[結(jié)果]1.實(shí)驗(yàn)組檢測結(jié)果:24例陽性,4例陰性;對照組培養(yǎng)結(jié)果:17例陽性,11例陰性。實(shí)驗(yàn)組檢測結(jié)果為陽性的24例標(biāo)本中有7例行細(xì)菌分離培養(yǎng)結(jié)果為陰性;實(shí)驗(yàn)組中檢測結(jié)果為陰性的4例標(biāo)本,行細(xì)菌分離培養(yǎng)同樣為陰性。2.實(shí)驗(yàn)組耗時(shí)約2h,對照組耗時(shí)約4-7d。3.實(shí)驗(yàn)組檢測出的病原菌以金黃色葡萄球菌最多見(11株,39.3%),其次是大腸埃希菌(5株,18%)、耐甲氧西林葡萄球菌(3株,10.7%)、結(jié)核桿菌(3株,10.7%),肺炎克雷伯菌(2株,7.1%)、銅綠假單胞菌(2株,7.1%),鮑曼不動(dòng)桿菌(2株,7.1%)。對照組培養(yǎng)出的病原菌以金黃色葡萄球菌最多見(8株,40%),其次是大腸埃希菌(5株,25%)、耐甲氧西林葡萄球菌(3株,15%)、肺炎克雷伯菌(2株,10%)、銅綠假單胞菌(1株,5%),鮑曼不動(dòng)桿菌(1株,5%)。4.實(shí)驗(yàn)組恒溫?cái)U(kuò)增微控流芯片核酸分析技術(shù)檢測陽性率為85.7%(24/28),高于對照組細(xì)菌培養(yǎng)陽性率60.7%(17/28),差異有統(tǒng)計(jì)學(xué)意義(χ2=4.462,p0.05)。Kappa 值為 0.66,一致性較好。[結(jié)論]1.恒溫?cái)U(kuò)增微控流芯片核酸分析技術(shù)檢測人工關(guān)節(jié)置換術(shù)后感染病原菌與細(xì)菌分離培養(yǎng)相比有較高的陽性檢出率,可有效的避免傳統(tǒng)細(xì)菌分離培養(yǎng)結(jié)果的假陰性。并且在耗時(shí)和效率方面較傳統(tǒng)的細(xì)菌分離培養(yǎng)相比具有明顯的優(yōu)勢。2.資料中,人工關(guān)節(jié)置換術(shù)后假體周圍感染病原菌以金黃色葡萄球菌等革蘭氏陽性菌為主,其次是革蘭氏陰性菌。3.本課題研究中,恒溫?cái)U(kuò)增微控流芯片核酸分析技術(shù)對于結(jié)核桿菌的檢測較細(xì)菌分離培養(yǎng)相比有一定的優(yōu)勢,顯示了良好的應(yīng)用前景。綜上所述,恒溫?cái)U(kuò)增微控流芯片核酸分析技術(shù)可適用于人工關(guān)節(jié)置換術(shù)后假體周圍感染病原菌的檢測,并具有便捷、快速及高陽性檢出率,可早期診斷病原菌、指導(dǎo)有效治療,從而縮短了抗感染治療的時(shí)間,增進(jìn)了患者的快速康復(fù),因此,具有較高的臨床推廣價(jià)值。
[Abstract]:Infection after artificial joint replacement (Artificial joint replacement, AJR) is one of the most serious complications after artificial joint replacement. It is an unavoidable problem for Joint Surgeons. The key to diagnosis is to find pathogenic bacteria. Once the pathogenic bacteria are identified, the diagnosis can be determined and the corresponding treatment can be started. The methods used to detect pathogenic bacteria are bacteria isolation and culture, long time consuming, and many factors that cause false negative results easily lead to the best time to delay the best treatment. Speed, high sensitivity, high accuracy, simple and efficient and so on. It discarded the shortcomings of the traditional polymerase chain reaction (polymerase chain reaction, PCR) technique with tedious operation, strict temperature requirement, and easy to pollute the specimen. From the beginning of the specimen treatment to the detection result, the whole course only needs 2 small. At present, the technique of chip nucleic acid analysis is mainly used for the detection of respiratory pathogenic bacteria. In 2518 samples of 5 large three A hospitals in China, the accuracy is up to 94.5%. because this technique is a new technology in the past two years. It has not been applied to the detection of pathogenic bacteria after AJR, and there is no related literature at home and abroad. The effect of nucleic acid analysis on microcontrolled flow microchip nucleic acid analysis in the detection of pathogenic bacteria after AJR was studied. The clinical practicality and popularization value of this technique were discussed. With the development of science and technology and the close combination of clinical and scientific research, the technology of constant temperature amplification must have one seat in the diagnosis of pathogenic bacteria. [Objective] [Objective] to explore the clinical practicality and popularization value by analyzing the effect of Nucleic Acid Analysy Technique of Isothermal Amplification Microfluidic Chip, NAATIAMC for rapid detection of pathogenic bacteria after AJR operation, and to explore the value of its clinical practicability and popularization. [square method]1.] selected Kunming Medical University from January 2016 to February 2017. In the Department of orthopedics and Chenggong branch of the First Affiliated Hospital of the First Affiliated Hospital, 28 patients with suspected artificial joint replacement were studied, including 12 male, 16 female, 64.5 + 13.1 years old (37~83 years old), 8 cases after artificial knee replacement and 20 cases of.2. after artificial hip replacement. The puncture fluid was divided into two groups. The specimens were divided into two groups, each of which was taken by the constant temperature amplification microcontrol flow chip nucleic acid analysis technique (experimental group) and the bacterial isolation and culture method (control group). The pathogen detection / culture.3. records were recorded in the experimental group and the control group. The species and time consuming of the pathogens were detected / cultured, and the pathogen detection was listed. The results of the results were compared between the two groups and the consistency of the results of the four groups was tested by the hypothesis of consistency. The positive rates of the two groups were compared with the x 2 test, P0.05, indicating that the difference was statistically significant. [results the test results of the]1. experimental group, 24 cases positive, 4 negative, 17 cases positive and 11 negative. The test group was positive 2. The results of bacterial isolation and culture were negative in 7 of the 4 specimens. In the experimental group, 4 specimens with negative test results, bacterial isolation and culture were same as 2h in the negative.2. experimental group, and the control group took the time of 4-7d.3. test to detect Staphylococcus aureus most (11, 39.3%), followed by Escherichia coli (5, 18%). Methicillin resistant Staphylococcus (3 strains, 10.7%), Mycobacterium tuberculosis (3 strains, 10.7%), Klebsiella pneumoniae (2 strains, 7.1%), Pseudomonas aeruginosa (2 strains, 7.1%), Acinetobacter Bauman (2, 7.1%). The most common bacteria in the control group were Staphylococcus aureus (8, 40%), followed by Escherichia coli (5, 25%), methicillin resistant Staphylococcus. 15%) Klebsiella pneumoniae (2 strains, 10%), Pseudomonas aeruginosa (1 strains, 5%), Acinetobacter Bauman (1, 5%).4. experimental group with constant temperature amplification microcontrolled flow chip nucleic acid analysis technique, the positive rate was 85.7% (24/28), higher than the control group of bacterial culture positive rate 60.7% (17 /28), the difference was statistically significant (chi 2=4.462, P0.05).Kappa value was 0.66, conformance comparison [conclusion]1. isothermal amplification microcontrolled flow chip nucleic acid analysis technology has a higher positive rate compared with bacteria isolation and culture after artificial joint replacement, which can effectively avoid false negative of traditional bacterial isolation and culture results, and is more obvious in time and efficiency than traditional bacterial isolation and culture. In the predominant.2. data, the pathogenic bacteria around the prosthesis were mainly Gram-positive bacteria such as Staphylococcus aureus, followed by the Gram-negative bacteria.3., and the nucleic acid analysis technology of the isothermal amplification micro control flow chip has a certain advantage compared with the bacterial isolation and culture. In summary, the nucleic acid analysis technology of isothermal amplification micro control flow chip can be applied to the detection of pathogenic bacteria around the prosthesis after artificial joint replacement, and it has the convenience, rapid and high positive detection rate. It can diagnose the pathogenic bacteria early and guide effective treatment, thus shortening the time of anti infection treatment and promoting the patient. Therefore, it has high clinical value.
【學(xué)位授予單位】：昆明醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R687.4

【參考文獻(xiàn)】

相關(guān)期刊論文 前10條

1 袁俊;馮建民;;人工關(guān)節(jié)置換術(shù)后假體周圍感染的病原學(xué)診斷研究進(jìn)展[J];中華關(guān)節(jié)外科雜志(電子版);2016年04期

2 王常德;康鵬德;裴福興;沈彬;周宗科;楊靜;;全關(guān)節(jié)置換術(shù)后假體周圍感染診斷的研究進(jìn)展[J];中國矯形外科雜志;2016年10期

3 吐爾洪江·瓦哈甫;沈?yàn)?張先龍;;表皮葡萄球菌引起假體周圍感染的生物膜形成機(jī)制[J];國際骨科學(xué)雜志;2016年02期

4 趙新;侯曉紅;;核酸恒溫?cái)U(kuò)增檢測技術(shù)發(fā)展態(tài)勢分析[J];基因組學(xué)與應(yīng)用生物學(xué);2016年03期

5 劉康;趙建寧;;人工關(guān)節(jié)置換術(shù)后假體周圍感染診斷的研究進(jìn)展[J];醫(yī)學(xué)研究生學(xué)報(bào);2015年09期

6 高川渤;袁紹輝;;人工關(guān)節(jié)置換術(shù)后感染病原菌譜的臨床意義[J];中國骨與關(guān)節(jié)雜志;2015年04期

7 周珍文;;分枝桿菌的實(shí)驗(yàn)室診斷[J];中華檢驗(yàn)醫(yī)學(xué)雜志;2014年03期

8 蔣梅花;陸勇;;人工關(guān)節(jié)置換術(shù)后并發(fā)癥影像學(xué)評價(jià)[J];國際骨科學(xué)雜志;2013年05期

9 羅英;;磁珠法核酸自動(dòng)提取儀在分子生物學(xué)領(lǐng)域的應(yīng)用[J];蠶學(xué)通訊;2013年02期

10 杜少華;黃強(qiáng)開;白潔玉;程鵬;梁大川;楊自權(quán);;關(guān)節(jié)置換術(shù)后假體周圍感染的診斷[J];醫(yī)學(xué)綜述;2012年14期

，

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