發(fā)布時間：2018-06-03 02:19

  本文選題：丙型肝炎病毒 + 檢測策略�。� 參考：《中國疾病預(yù)防控制中心》2014年碩士論文

【摘要】：研究背景 丙型肝炎病毒(Hepatitis C virus, HCV)主要通過血液傳播,引起的丙型病毒性肝炎(以下簡稱“丙型肝炎”)呈世界范圍流行。由于丙型肝炎發(fā)病隱匿、癥狀不典型,因此其臨床診斷主要依靠實驗室的檢測結(jié)果。HCV特異性抗體是丙型肝炎臨床診斷中實驗室檢測的主要指標,針對HCV抗體的檢測主要分為篩查試驗和補充試驗兩部分�？贵w的篩查試驗,篩查試驗一般采用酶聯(lián)免疫吸附法(Enzyme Linked Immunosorbent Assay, ELISA),對于篩查實驗結(jié)果呈陽性反應(yīng)的樣本采用特異性更好的補充試驗對結(jié)果進行確證,補充實驗通常采用重組免疫印跡試驗(Recombinant Immunoblot Assay,RIBA),篩查實驗和補充實驗配合使用,形成一系列的檢測策略,從而為臨床診斷提供可靠的結(jié)果。 我國各級臨床實驗室雖然已經(jīng)廣泛開展了HCV相關(guān)檢測,但是仍然沒有明確的檢測策略以及對應(yīng)的結(jié)果解釋。對此,衛(wèi)生與計劃生育委員會委托中國疾控中心編寫了《丙型肝炎病毒實驗室檢測技術(shù)規(guī)范》(試行)(以下簡稱《規(guī)范》),其中對HCV臨床診斷的檢測策略做了詳細的要求,提出了HCV臨床診斷抗體篩查策略、HCV臨床診斷核酸檢測策略等。HCV臨床診斷核酸檢測策略是對HCV抗體篩查陽性反應(yīng)的樣本采用核酸擴增檢(Nucleric acid Amplification Test, NAT)和RIBA組合使用的檢測方法。任何一種疾病檢測策略在應(yīng)用之前,都必須經(jīng)過嚴密的評估,確定其可用性、準確性、可靠性后方可推廣使用。我國新頒發(fā)的《規(guī)范》中規(guī)定的臨床診斷實驗室檢測策略己完成了抗體檢測策略的評估,本研究主要圍繞核酸檢測策略部分進行評估。 研究目的 1.對后續(xù)將用于HCV臨床i診斷核酸檢測策略中的HCV核酸檢測試劑的性能進行評估。 2.在國家艾滋病參比實驗室進行HCV臨床診斷核酸檢測策略的實驗室小樣本范圍評估。 3.在多個現(xiàn)場試點進行HCV臨床診斷核酸檢測策略應(yīng)用的評估,進行證明性研究,提供一種可推廣到全國多點使用的、合理的、可靠的檢測策略。 4.為國家級策略評估三階段中第三階段的HCV臨床診斷檢測策略實施后的評估奠定基礎(chǔ)。為進一步修訂《規(guī)范》中HCV檢測策略提供依據(jù),使我國丙型肝炎的臨床診斷實驗室檢測流程更規(guī)范。 研究方法 1.方法學評價部分：建立HCV核酸檢測基礎(chǔ)血清盤、HCV分亞型血清盤、HCV RNA線性血清盤、HCV RNA靈敏度血清盤、參比室干擾樣本血清盤、HCV核酸精密度血清盤。用HCV核酸檢測試劑對己建立的血清盤進行盲法檢測。對檢測結(jié)果進行計算求得待評估試劑的陽性符合率、陰性符合率、亞型檢出能力、線性、分析靈敏度、分析特異性、精密度等指標以評價檢測方法的總體性能。 2.臨床診斷核酸檢測策略評估部分的研究包括兩個階段：第一階段,建立一套本底信息明確的血清盤用于HCV臨床診斷核酸檢測策略在國家艾滋病參比實驗室范圍內(nèi)的評估。第二階段,選擇北京、天津兩地作為試點,對所選的試點現(xiàn)場進行丙型肝炎臨床診斷核酸檢測策略的大樣本評估,參與評估的樣本分別來自2013年HCV監(jiān)測哨點和HIV監(jiān)測哨點的吸毒人群以及北京市腎透析丙型肝炎流行病學調(diào)查的腎透析人群。兩個評估階段均與抗體檢測補充策略即RIBA單獨作為篩查陽性樣本補充實驗的檢測策略相比較,計算檢測策略的符合率等指標,評估檢測策略的應(yīng)用效果,同時對核酸檢測策略在不同人群、不同地點、不同樣本量檢測的總體性能進行比較。 結(jié)果 1. HCV RNA檢測試劑的陽性符合率為49/50(98%)；陰性符合率為50/50(100%),總符合率為99/100(99%)。對各種可能對臨床檢測產(chǎn)生干擾的樣本的分析特異性為100%。對HCV1到6個型別共9種亞型均具有檢出能力。對HCV RNA濃度分別為低、中、高(102、104、106IU/mL)三份樣本的檢測中,除低濃度樣本外對中、高濃度樣本的天內(nèi)精密度均小于6%,天間精密度分別為9.28%、5.03%、1.53%,總精密度分別為11.55%、4.45%、3.08%。磁珠提取HCV核酸檢測試劑與Roche COBAS AmpliPrep/COBAS TaqMan HCV Test定量結(jié)果間的線性相關(guān)系數(shù)r值為0.9335。 2.HCV臨床診斷核酸檢測策略在參比室小樣本及現(xiàn)場大量樣本進行應(yīng)用的檢測敏感性、特異性、陽性預(yù)測值、陰性預(yù)測值均能達到100%。HCV臨床診斷核酸檢測策略的整體檢測性能在吸毒人群和腎透析人群中無統(tǒng)計學差異(P0.05)。策略的整體檢測性能在應(yīng)用于參比實驗室的小樣本與試點現(xiàn)場大樣本之間有強一致性,兩者之間無統(tǒng)計學差異。 結(jié)論 1.HCV核酸檢測試劑有較高的陽性符合率和陰性符合率,且精密度、靈敏度都較高,對各亞型均由檢出能力,適合用于HCV臨床診斷核酸檢測策略的評估。 2.我國《丙型肝炎病毒實驗室檢測技術(shù)規(guī)范》(試行)中的臨床診斷核酸檢測策略在參比實驗室的小范圍內(nèi)及多個試點現(xiàn)場的應(yīng)用效果均較好,對不同HCV感染率人群的檢測效力差異不明顯,具有普遍適用性,可有效降低檢測的假陽性率,并顯著降低檢測成本。
[Abstract]:Research background
Hepatitis C virus (Hepatitis C virus, HCV) is mainly transmitted through blood, causing hepatitis C virus hepatitis (hereinafter referred to as "hepatitis C") in the world. Due to HCV occult, symptoms are not typical, so its clinical diagnosis mainly relies on laboratory test results of.HCV specific antibody is the clinical diagnosis of hepatitis C The main index of detection in the laboratory of HCV is divided into two parts: screening test and supplementary test. Screening test of antibody, screening test is usually used in Enzyme Linked Immunosorbent Assay, ELISA, and it is better to supplement the samples with positive reaction in screening experiment. The results were confirmed by the experiment. The supplementary experiment was usually used in combination with the recombinant immunoblotting test (Recombinant Immunoblot Assay, RIBA), the screening experiment and the supplemental experiment, and a series of detection strategies were formed to provide reliable results for clinical diagnosis.
Although the clinical laboratory at all levels in our country has widely carried out HCV related tests, there is still no clear detection strategy and corresponding interpretation. In this regard, the health and Family Planning Commission has commissioned the China CDC to compile a "hepatitis C virus laboratory test specification" (hereinafter referred to as < standard >), of which HCV The detection strategy of clinical diagnosis has been required in detail. The HCV clinical diagnostic antibody screening strategy, the HCV clinical diagnostic nucleic acid detection strategy, and.HCV clinical diagnostic nucleic acid detection strategy are the detection methods used for the combination of nucleic acid amplification test (Nucleric acid Amplification Test, NAT) and RIBA for the positive reaction of HCV antibody screening. Before application, any kind of disease detection strategy must be carefully evaluated to determine its availability, accuracy and reliability. The clinical diagnosis laboratory testing strategy stipulated in the newly issued < standard > has completed the evaluation of antibody detection strategy. This research focuses on the part of nucleic acid detection strategies. Assessment.
research objective
1. evaluate the performance of the HCV nucleic acid reagents that will be used in the clinical I diagnostic nucleic acid detection strategy of HCV.
2. in the National AIDS reference laboratory, a small sample size assessment of HCV clinical diagnostic nucleic acid detection strategy was conducted.
3. the evaluation of the application of the HCV clinical diagnostic nucleic acid detection strategy was conducted in several field trials, and a demonstration study was carried out to provide a reasonable and reliable detection strategy that could be extended to the national multipoint use.
4. lay the foundation for the assessment of the implementation of the third stage HCV clinical diagnosis strategy in the three stage of the national strategy assessment, and to provide a basis for further revising the HCV detection strategy in the < standard > to make the laboratory testing flow of hepatitis C in China more standardized.
research method
The 1. methodology evaluation part: establish HCV nucleic acid test basal blood disks, HCV subtype blood disks, HCV RNA linear sera disc, HCV RNA sensitivity sera disc, reference room interfering sample sera disc, HCV nucleic acid precision blood disks. The serum disk entry blind method was detected by HCV nucleic acid detection reagent. The test results were calculated and evaluated. The positive coincidence rate, negative coincidence rate, subtype detection ability, linearity, analysis sensitivity, specificity and precision were used to evaluate the overall performance of the method.
2. the study of the 2. clinical diagnostic nucleic acid detection strategy includes two stages: the first stage, the establishment of a set of background information clear serum plates for the HCV clinical diagnostic nucleic acid detection strategy in the National AIDS reference laboratory assessment. The second stage, the choice of Beijing, Tianjin as a pilot, the selected pilot site into the site. A large sample assessment of nucleic acid detection strategies for the clinical diagnosis of hepatitis C was conducted, and the samples participated in the assessment were from the drug users in the HCV sentinel and HIV sentinel points in 2013 and the renal dialysis population in the epidemiological investigation of HCV in Beijing. The two assessment stages were all screened by the anti physical examination supplementation strategy, RIBA alone. The test strategy of positive samples was compared, the coincidence rate of the detection strategy was calculated, and the application effect of the detection strategy was evaluated. At the same time, the overall performance of the detection strategy of nucleic acid detection strategy in different population, different locations and different sample quantities was compared.
Result
The positive coincidence rate of the 1. HCV RNA detection reagents was 49/50 (98%); the negative coincidence rate was 50/50 (100%) and the total coincidence rate was 99/100 (99%). The specificity of the analytical specificity for various possible samples that could interfere with clinical detection was detected by 100%. on 9 subtypes from HCV1 to 6 types. The RNA concentration of HCV was low, medium and high (102104106IU/mL), respectively. In the detection of three samples, in addition to the low concentration samples, the precision of the high concentration samples was less than 6%, the day precision was 9.28%, 5.03%, 1.53%, respectively, the total precision was 11.55%, 4.45%, 3.08%. magnetic beads extracted HCV nucleic acid detection reagent and the linear correlation coefficient r value between Roche COBAS AmpliPrep/ COBAS TaqMan HCV Test quantitative results. For 0.9335.
The detection sensitivity, specificity, positive predictive value and negative predictive value of the 2.HCV clinical diagnostic nucleic acid detection strategy used in the small sample of the reference room and the large number of samples in the field can reach the overall detection performance of the 100%.HCV clinical diagnostic nucleic acid detection strategy. There is no statistical difference between the drug addicts and the renal dialysis population (P0.05). There was no statistical difference in the detection performance between the small samples used in the reference laboratory and the large sample sites at the pilot site.
conclusion
The 1.HCV nucleic acid detection reagent has high positive coincidence rate and negative coincidence rate, and the precision is high, and the sensitivity is high. All the subtypes are detected. It is suitable for the evaluation of nucleic acid detection strategy in the clinical diagnosis of HCV.
2. the clinical diagnostic nucleic acid detection strategy in the laboratory test of hepatitis C virus (HCV) in China has better application effect in the small range of the reference laboratory and in several pilot sites. The difference of detection effectiveness for people with different HCV infection rates is not obvious, and it is universally applicable, which can effectively reduce the false positive rate of detection. Significantly reduce the cost of detection.
【學位授予單位】：中國疾病預(yù)防控制中心
【學位級別】：碩士
【學位授予年份】：2014
【分類號】：R512.63

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