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1.Fever pathogen

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

2.Infection of pathogens

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.Respiratory pathogens

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

4.Intestinal pathogens

4.1Applicability of duplex real time and lateral flow stripreverse-transcription recombinase aided amplifica-tion assays for the detection of Enterovirus 71 andCoxsackievirus A16

4.2Development of a reverse transcription recombi-nase-aided amplifcation assay for the detection ofcoxsackievirus A10 and coxsackievirus A6 RNA

5.Disease surveillance

5.1Development and evaluation of a rapid detectionassay for severe fever with thrombocytopeniasyndrome virus based on reverse-transcriptionrecombinase polymerase amplification

5.2lntegrating Microwave Resonator and Microchannelwith Immunochromatographic Strip for Stable andQuantitative Biodetection

5.3lnternally 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

5.4A 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).