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1.發(fā)熱病原體

1.1Rapid visual detection of dengue virus by combiningreverse transcription recombinase-aided amplifica-tion with lateral-flow dipstick assay

1.2A Reverse-transcription Recombinase-aided Amplifi-cation Assay for the Rapid Detection of the Far-East-ern Subtype of Tick-borne Encephalitis Virus

1.3Development of an lnternally Controlled ReverseTranscription Recombinase-aided Amplification Assayfor the Rapid and Visual Detection of West Nile Virus

1.4重組酶介導擴增方法快速檢測黃熱病毒

1.5逆轉(zhuǎn)錄重組酶介導擴增技術(shù)快速檢測基孔肯雅熱病毒

1.6實時熒光逆轉(zhuǎn)錄重組酶介導的等溫擴增技術(shù)檢測寨卡病毒方法的建立

1.7西尼羅病毒的逆轉(zhuǎn)錄重組酶介導擴增檢測方法

1.8雙重熒光RT-RAA檢測5種蚊媒病毒方法的建立

1.9塔希納病毒的重組酶介導擴增快速檢測方法

2.感染類病原體

2.1A rapid and sensitive recombinase aided amplificationassay to detect hepatitis Bvirus without DNAextraction

2.2Field applicable detection of hepatitis B virus usinginternal controlled duplex recombinase-aided ampli-fication assay and lateral flow dipstick assay

3.呼吸道病原體

3.1Multiple-centre clinical evaluation of an ultrafastsingle-tube assay for SARS-CoV-2 RNA

3.2A Reverse-Transcription Recombinase-Aided Amplifi-cation Assay for Rapid Detection of the 2019 NovelCoronavirus (SARS-CoV-2)

3.3A multi-country phase 2 study to evaluate the suit-case lab for rapid detection of SARS-CoV-2 in sevenSub-Saharan African countries: Lessons from the field

3.4Use of a rapid reverse-transcription recom binaseaided amplification assay for respira tory syncytialvirus detection

3.5Development of a duplex reverse transcriptionrecombinase-aided amplifcation assay for respiratorysyncytial virus incorporating an internal control

3.6A rapid and sensitive recombinase aided amplificationassay incorporating competitive internal control todetect Bordetella pertussis using the DNA obtained by boiling

3.7Use of a rapid recombinase-aided amplification assayfor Mycoplasma pneumoniae detection

3.8Development and evaluation of recombinase-aidedamplification assays incorporating competitive intenal controls for detection of human adenovirusserotypes 3 and 7

3.9中東呼吸綜合征冠狀病毒重組酶介導核酸檢測方法的建立

3.10中東呼吸綜合征冠狀病毒RT_RAA快速檢測方法的建立及應(yīng)用

3.11甲型流感病毒的逆轉(zhuǎn)錄重組酶介導核酸擴增快速檢測方法研究

3.12重組酶介導恒溫擴增快速檢測人鼻病毒的方法學研究

3.13新型冠狀病毒肺炎核酸檢測和診斷專利信息分析

3.14新型冠狀病毒核酸熒光型RT-RAA檢測方法的建立及其評價

3.15基于RAA熒光法快速檢測呼吸道腺病毒的研究

4.分枝桿菌病原體

4.1應(yīng)用重組酶介導的恒溫擴增法建立檢測非結(jié)核分枝桿菌的方法研究

4.2探針導向重組酶介導等溫擴增法檢測TB_rpoB基因突變的應(yīng)用價值

4.3RNA恒溫擴增實時檢測技術(shù)與熒光定量PCR聯(lián)合檢測肺泡灌洗液對痰涂陰性肺結(jié)核的快速診斷價值

5.腸道病原體

5.1Applicability of duplex real time and lateral flow stripreverse-transcription recombinase aided amplifica-tion assays for the detection of Enterovirus 71 andCoxsackievirus A16

5.2Development of a reverse transcription recombi-nase-aided amplifcation assay for the detection ofcoxsackievirus A10 and coxsackievirus A6 RNA

5.3RAA聯(lián)合CRISPR-Cas13a快速檢測4種腹瀉病原菌

6.疾病監(jiān)測

6.1Development and evaluation of a rapid detectionassay for severe fever with thrombocytopeniasyndrome virus based on reverse-transcriptionrecombinase polymerase amplification

6.2lntegrating Microwave Resonator and Microchannelwith Immunochromatographic Strip for Stable andQuantitative Biodetection

6.3lnternally controlled recombinase-aided amplifcation(IC-RAA) assays for the detection of human papillo-mavirus genotypes 16 and 18 using extracted DNAand samples treated with nucleic acid releasing agent

6.4A probe directed recombinase amplification assay fordetection of MTHER A1298C polymor phis associatedwith congenital heart disease

Rapid visual detection of dengue virus bycombining reverse transcription recombi-nase-aided amplification with lateral-flowdipstick assay

Abstract:Objectives:Dengue caused by infection with the dengue virus (DENV) is endemic in the tropical andsubtropical regions of the world and of greatest public health concern.With more large outbreaks in rural areas,the purpose of this study was to develop a point-of-care test using recombinase-aided amplification and later-al-flow dipsticks for rapidly detecting DENV in low-resource settings.Methods:The primers for the recombi-nase-aided amplification (RAA) assay were designed based on 3'UTR of the DENV genome and screened.TheRAA temperature,time and the concentration of primers were thenoptimized,as well as thelateral-flow dipstickassay (LFD) time.Finally,the diagnostic performance of the reverse transcription(RT)-RAA-LFD assay wasevaluated using blood samples from 247 patients who were clinically suspected to be infected with DENV.Re-sults:The RAA primer pair F1/R2 was the optimal combination for detecting DENV.The RT-RAA was performedin an incubator block at 37℃ for 20 minutes,and the amplicons were visible in the flow dipsticks from a nakedeye within 3 minutes.The detection limit of the developed RT-RAA-LFD assay was 10 copies/mL with high spec-ificity for DENV.Compared with commercial reverse transcription quantitative PCR assay,the kappa value ofRT-RAA-LFD in the 247 clinical samples was 0.957.Conclusions:In this study,a rapid and visual point-of-caretest based on RT-RAA and LFD assay was developed. It was found to be suitable for reliable detection of DENVin low-resource settings with limited aboratory capabilities and optimal storage conditions.

Keywords:Dengue virus Recombinase-aided amplification Lateral-flow dipstick assay.

A Reverse-transcription Recombinase-aidedAmplification Assay for the Rapid Detection ofthe Far-Eastern Subtype of Tick-borne En-cephalitis Virus

Abstract: Objective Tick-borne encep halitis virus(TBEV) is an emerging pathogen in Europe and North Asiathat causes tick-borne encephalitis(TBE).A simple,rapid method for detecting TBEV RNA is needed to controlthis disease.Methods Areverse-transcription recombinase-aided amplification(RT-RAA) assay was deseloped.This assay can be completed in one elosed tube at 39 ℃ within 30 minutes.The sensitivity and specificity ofRT-RAA were validated using non-infectious synthetic RNA represerting a fragment of the NS5 region of thewild-type(WT) TBEV genome and the Senzhang strsain.Additionally.10 batches of tick samples were used toevaluate the performance of the RT-RAA ass8ty.Resudts The analyticalimit of detection of the assay was 20copies per reaction of the TBEV syrthetic transcript and 3 plsque-forming units (pfu) per resction of TBEVtiters.With the specific assay,no signal due to other ar boviruses was observed.Of the 10 batches of tick.samplesobtained from the Changbai Mountains of Chins, three were TBEV-positive,which was consistent withthe results of the quantitative real-time PCR assay.Conclusion A rapid,highly sensitive,specific,andeasy-to-use method was developed for the detection of the TBEV Far-Eastern subtype.

Key words: Tick-borne encephalitis virus;Subtype;Far-eastern;Detection;RT-RAA.

Development of an Internally Controlled Re-verse Transcription Recombinase-aided Am-plification Assayfor the Rapid and VisualDetection of West Nile Virus

West Nile virus wNV causes West Nile feverand West Nile encephalitis.Because irnfection by Wv cresatesserisus public health problems,its simple,rapid,and visual dietection is very important in clirical practice,espe-cially in resource-lmited laboratories We hase developed a rapid,specific,arnd highly senitive internallycontrdlled reverse transription recombinase-aided amplific ation(RT RAA) assay to detect wNv,using bothresl-time flucrescence and the bteral flow dinstick (LFD) at 390 "C for 30 mn.The analyfical sersitivity ofthe RT-RAA ass8ay was 10 plasmid copis and 1.6 pfuper reaction with real-time fhuorescence,and 1p00plasmidcopiespefreaction wih theLFD.Nocross reaction with other contrd viruses was observed.Compared with theRT-qPCR assay.the RT-RAA assay dermonstrated 100% sensitivity and 100% specificity for WIw.

Key words:WrNV;Detectior RT-RAA;Lateral flow dipstick (LFD).

重組酶介導擴增方法快速檢測黃熱病毒

摘要：目的本研究采用重組酶介導的等溫核酸擴增方法(RAA)，通過使用逆轉(zhuǎn)錄酶，建立黃熱病毒的一步法等溫核酸擴增(RT一RAA)方法。方法根據(jù)黃熱病毒基因組保守序列設(shè)計引物和探針，建立并分析RT一RAA的重復性、特異性、靈敏度；所以建立方法對黃熱病毒樣本進行檢測，同時以基因測序進行驗證。結(jié)果黃熱病毒RT一RAA擴增，體系中加入40U的逆轉(zhuǎn)錄酶擴增效果最佳。該方法檢測時間短(<20min)。并且靈敏度高，檢測下限可達100copy，與登革病毒、西尼羅病毒、日本乙型腦炎病毒、基孔肯雅熱病毒等蚊媒病毒無交叉反應(yīng)，具有良好的特異性。結(jié)論――構(gòu)建的黃熱病毒RT一RAA方法具有快速，特異以及靈敏的特點,適應(yīng)于黃熱病毒的口岸快速檢測。

關(guān)鍵詞 : 重組酶介導擴增；黃熱病毒；分子檢測。