本文選題：瘧疾　切入點(diǎn)：鏡檢　出處：《安徽醫(yī)科大學(xué)》2014年碩士論文　論文類型：學(xué)位論文

【摘要】：目的采用鏡檢、RDT和PCR三種檢測方法對(duì)網(wǎng)報(bào)瘧疾病例復(fù)核檢測，比較三種方法在瘧疾診斷中的敏感性、特異性，為科學(xué)地分析判斷復(fù)核檢測結(jié)果，以及RDT能否在基層替代瘧原蟲鏡檢提供依據(jù)。RDT檢測時(shí)，同時(shí)使用進(jìn)口和國產(chǎn)RDT兩種產(chǎn)品進(jìn)行檢測，通過比較兩種產(chǎn)品的敏感性、特異性，為監(jiān)測工作中產(chǎn)品選擇的可行性提供依據(jù)。動(dòng)態(tài)觀察瘧疾病例鏡檢、PCR和RDT的檢測陰轉(zhuǎn)時(shí)間，以及經(jīng)治療后瘧疾病人鏡檢原蟲的密度隨時(shí)間的變化，為瘧疾監(jiān)測和防治工作提供科學(xué)的理論依據(jù)。 方法收集安徽省2012年1月~2013年9月中國疾病預(yù)防控制信息傳染病報(bào)告系統(tǒng)的網(wǎng)報(bào)267例瘧疾病例的血片、抗凝血，病例報(bào)告類型包括實(shí)驗(yàn)室、臨床和疑似三種；50例非瘧疾病人血樣，來自芙蓉社區(qū)衛(wèi)生服務(wù)中心。267例流行病學(xué)調(diào)查表來自寄生蟲病防治信息管理系統(tǒng)。對(duì)血片和抗凝血分別進(jìn)行鏡檢、RDT和PCR檢測，，并將快速檢測試劑條的結(jié)果與鏡檢法和PCR的結(jié)果對(duì)比分析其敏感性與特異性。收集安徽省立醫(yī)院及肥東縣人民醫(yī)院2013年5月~2013年9月瘧疾住院病人37例，確診后連續(xù)采血7天，若病人治愈出院、轉(zhuǎn)院等情況，以鏡檢與PCR結(jié)果有轉(zhuǎn)陰時(shí)間為標(biāo)準(zhǔn)，納入研究范圍。使用EDTA管采集血樣，并對(duì)病人進(jìn)行流行病學(xué)調(diào)查。每天采血后立即涂制血片，同時(shí)做RDT，并提取DNA準(zhǔn)備做PCR。采用三種方法進(jìn)行檢測，觀察不同檢測方法的陰轉(zhuǎn)時(shí)間，經(jīng)SPSS11.5軟件統(tǒng)計(jì)分析，采用非參數(shù)檢驗(yàn)（nonparametric tests）中的Wilcoxon檢驗(yàn)比較，P＜0.05認(rèn)為差異有統(tǒng)計(jì)學(xué)意義。 結(jié)果收集267例網(wǎng)報(bào)病例的血片，鏡檢復(fù)核瘧原蟲陽性148例，陽性率為55.43%，其中惡性瘧117例，間日瘧20例，卵形瘧10例，三日瘧1例；267例抗凝血樣采用PCR法檢測，瘧原蟲陽性者150例，陽性率為56.18%，其中惡性瘧119例（包括鏡檢檢出的117例），間日瘧20例，卵形瘧10例，三日瘧1例；采用進(jìn)口RDT試劑對(duì)267份抗凝血血樣進(jìn)行檢測，檢測出147例陽性，陽性率為55.06%，其中非惡性瘧21例（包括鏡檢和PCR均檢為間日瘧的19例和鏡檢和PCR均檢為惡性瘧的2例），惡性瘧126例（包括鏡檢為惡性瘧的114例，另12例鏡檢陰性；PCR檢為惡性瘧116例，另10例PCR檢為陰性），因此，RDT未檢出1例間日瘧和1例惡性瘧，并將2例惡性瘧檢為非惡性瘧，10例PCR和鏡檢均檢為陰性的病例檢為惡性瘧，10例卵形瘧和1例三日瘧均未檢出；50例非瘧疾病人RDT、鏡檢和PCR檢測均為陰性。以鏡檢為標(biāo)準(zhǔn)，對(duì)間日瘧和惡性瘧進(jìn)行分析得出，RDT的敏感度為98%，特異度90%，假陽性率10%，假陰性率2%，陽性預(yù)測值92%，陰性預(yù)測值98%；以PCR為標(biāo)準(zhǔn)，對(duì)間日瘧和惡性瘧進(jìn)行分析得出，RDT的敏感度為98%，特異度91%，假陽性率9%，假陰性率2%，陽性預(yù)測值93%，陰性預(yù)測值98%。采用國產(chǎn)RDT檢測結(jié)果為陽性143例，陰性124例，其中非惡性瘧18例，惡性瘧125例（包括進(jìn)口RDT檢為惡性瘧的123例，另2例進(jìn)口RDT檢為非惡性瘧，但是鏡檢和PCR均檢為惡性瘧）。因此，國產(chǎn)RDT未檢出3例惡性瘧和1例間日瘧，卵形瘧和三日瘧均未檢出。將國產(chǎn)RDT的檢測結(jié)果與進(jìn)口RDT結(jié)果進(jìn)行比較，兩方法配對(duì)卡方檢驗(yàn)結(jié)果表明兩方法差異沒有統(tǒng)計(jì)學(xué)意義（2=4.000，P＞0.05），Kappa檢驗(yàn)結(jié)果P＜0.001，且Kappa值0.97，提示兩方法高度一致。動(dòng)態(tài)采集37例確診瘧疾病人血樣，鏡檢轉(zhuǎn)陰時(shí)間1~5天，中位數(shù)為2天；PCR轉(zhuǎn)陰時(shí)間1~6天，中位數(shù)為3天，經(jīng)非參數(shù)檢驗(yàn)（nonparametric tests）中的Wilcoxon檢驗(yàn)比較，差異有統(tǒng)計(jì)學(xué)意義（Z=3.776，P＜0.05），PCR轉(zhuǎn)陰時(shí)間約比鏡檢長1天；RDT的陰轉(zhuǎn)時(shí)間長于7天。37例病例鏡檢結(jié)果轉(zhuǎn)陰時(shí)間為1~5天不等，16例病人第二天即轉(zhuǎn)陰，其中1天后陰轉(zhuǎn)病例密度中位數(shù)為3571/μl血（267/μl血，38790/μl血），2~5天后陰轉(zhuǎn)病例密度中位數(shù)分別為第1天38385/μl血、第2天11456/μl血及第3天147/μl血，呈下降趨勢。 結(jié)論本研究結(jié)果表明，課題使用RDT產(chǎn)品無法檢出卵形瘧和三日瘧；對(duì)于惡性瘧和間日瘧，RDT與鏡檢和PCR相比，RDT的敏感性和特異性均較高。由于PCR在基層較難普及，而鏡檢對(duì)技術(shù)的要求較高而且人員培訓(xùn)周期長等原因，RDT有望在基層取代鏡檢進(jìn)行瘧疾監(jiān)測。國產(chǎn)RDT敏感性和特異性也較高，可以在基層取代進(jìn)口RDT進(jìn)行瘧疾病例監(jiān)測工作。三種方法RDT的有效檢測時(shí)間最長，長于7天，PCR次之，約為3天，鏡檢陰轉(zhuǎn)時(shí)間約為2天，PCR陰轉(zhuǎn)時(shí)間一般比鏡檢遲1天，為瘧疾病例復(fù)核以及疫點(diǎn)是否處置提供了依據(jù)。瘧疾病例原蟲密度變化可以作為臨床治療效果的判斷指標(biāo)。
[Abstract]:The purpose of the examination, three methods for detection of RDT and PCR on the network reported malaria cases review detection, sensitivity, comparison of the three methods in diagnosis of malaria specific, scientific analysis and review of test results, the RDT can replace blood smears at the grassroots level to provide the basis for.RDT detection, and the use of imported and domestic RDT detection of two kinds of products, through the comparison of two kinds of products, the sensitivity, specificity, and provide the basis for the feasibility of product selection and monitoring of malaria cases. Dynamic observation in microscope, detection of PCR and RDT negative conversion time, and the treatment of malaria parasite microscopic density changes with time, provide the scientific basis for malaria surveillance and control work.
Methods to collect blood, Anhui Province in January 2012 ~2013 year in September Chinese disease prevention and control report system of infectious disease information network reported 267 cases of malaria anticoagulant, case report types include laboratory, clinical and suspected three; 50 cases of non malaria patients blood samples from Furong community health service center.267 cases from the epidemiological survey of parasitic diseases the prevention and control of the information management system. The blood and anticoagulant were microscopy, RDT and PCR detection and rapid detection of contrast reagent strip results with microscopic examination and PCR analysis of the result of the sensitivity and specificity of the hospital and Anhui Provincial Hospital. Feidong County People's Hospital from May 2013 ~2013 year in September 37 cases of malaria patients, diagnosed after continuous blood 7 days, if the patient was cured and discharged, etc., to the microscopic examination and PCR negative time, included in the study. Blood samples were collected using EDTA tube, and the disease Human epidemiological survey was conducted every day. Blood immediately after coated with blood, do RDT at the same time, DNA was extracted by three methods for PCR. detection, observation of different detection methods of negative time is analyzed by SPSS11.5 software using non parametric test (nonparametric tests) compared with Wilcoxon test, P < 0.05 the difference was statistically significant.
緇撴灉鏀墮泦267渚嬬綉鎶ョ梾渚嬬殑琛

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