title: Development and Evaluation of a Rapid and Sensitive EBOV-RPA Test for Rapid Diagnosis of Ebola Virus Disease.
abstract: Confirming Ebola virus disease (EVD), a deadly infectious disease, requires real-time RT-PCR, which takes up to a few hours to yield results. Therefore, a rapid diagnostic assay is imperative for EVD diagnosis. A rapid nucleic acid test based on recombinase polymerase amplification (EBOV-RPA) was developed to specifically detect the 2014 outbreak strains. The EBOV-RPA assay was evaluated by testing samples from suspected EVD patients in parallel with RT-PCR. An EBOV-RPA, which could be completed in 20 min, was successfully developed. Of 271 patients who tested positive for Ebola virus by RT-PCR, 264 (sensitivity: 97%, 95% CI: 95.5-99.3%) were positive by EBOV-RPA; 101 of 104 patients (specificity: 97%, 95% CI: 93.9-100%) who tested negative by RT-PCR were also negative by EBOV-RPA. The sensitivity values for samples with a Ct value of <34, which accounted for 95.59% of the samples, was 100%. Discordant samples positive by RT-PCR but negative by EBOV-RPA had significantly high Ct values. Results of external quality assessment samples with EBOV-RPA were 100%, consistent with those of RT-PCR. The EBOV-RPA assay showed 97% sensitivity and 97% specificity for all EVD samples tested, making it a rapid and sensitive test for EVD diagnosis.
authors.affiliation: Institute of Disease Control and Prevention, Academy of Military Medical Sciences, 100071, Beijing, China.
authors.forename: Mingjuan
authors.initials: M
authors.lastname: Yang
authors.affiliation: Institute of Disease Control and Prevention, Academy of Military Medical Sciences, 100071, Beijing, China.|China Mobile Laboratory Response Team for Ebola in Sierra Leone, Freetown, Sierra Leone.
authors.forename: Yuehua
authors.initials: Y
authors.lastname: Ke
authors.affiliation: Institute of Disease Control and Prevention, Academy of Military Medical Sciences, 100071, Beijing, China.|China Mobile Laboratory Response Team for Ebola in Sierra Leone, Freetown, Sierra Leone.
authors.forename: Xuesong
authors.initials: X
authors.lastname: Wang
authors.affiliation: Institute of Disease Control and Prevention, Academy of Military Medical Sciences, 100071, Beijing, China.
authors.forename: Hang
authors.initials: H
authors.lastname: Ren
authors.affiliation: Institute of Disease Control and Prevention, Academy of Military Medical Sciences, 100071, Beijing, China.|China Mobile Laboratory Response Team for Ebola in Sierra Leone, Freetown, Sierra Leone.
authors.forename: Wei
authors.initials: W
authors.lastname: Liu
authors.affiliation: China Mobile Laboratory Response Team for Ebola in Sierra Leone, Freetown, Sierra Leone.|Key Laboratory of Jilin Province for Zoonosis Prevention and Control, Changchun 130122, China.
authors.forename: Huijun
authors.initials: H
authors.lastname: Lu
authors.affiliation: Institute of Disease Control and Prevention, Academy of Military Medical Sciences, 100071, Beijing, China.|China Mobile Laboratory Response Team for Ebola in Sierra Leone, Freetown, Sierra Leone.
authors.forename: Wenyi
authors.initials: W
authors.lastname: Zhang
authors.affiliation: Wangjing Hospital, China Academy of Traditional Chinese Medicine, 100102, Beijing, China.
authors.forename: Shiwei
authors.initials: S
authors.lastname: Liu
authors.affiliation: Institute of Disease Control and Prevention, Academy of Military Medical Sciences, 100071, Beijing, China.|China Mobile Laboratory Response Team for Ebola in Sierra Leone, Freetown, Sierra Leone.
authors.forename: Guohui
authors.initials: G
authors.lastname: Chang
authors.affiliation: Institute of Disease Control and Prevention, Academy of Military Medical Sciences, 100071, Beijing, China.|China Mobile Laboratory Response Team for Ebola in Sierra Leone, Freetown, Sierra Leone.
authors.forename: Shuguang
authors.initials: S
authors.lastname: Tian
authors.affiliation: China Mobile Laboratory Response Team for Ebola in Sierra Leone, Freetown, Sierra Leone.|Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China.
authors.forename: Lihua
authors.initials: L
authors.lastname: Wang
authors.affiliation: Institute of Disease Control and Prevention, Academy of Military Medical Sciences, 100071, Beijing, China.
authors.forename: Liuyu
authors.initials: L
authors.lastname: Huang
authors.affiliation: Institute of Disease Control and Prevention, Academy of Military Medical Sciences, 100071, Beijing, China.|China Mobile Laboratory Response Team for Ebola in Sierra Leone, Freetown, Sierra Leone.
authors.forename: Chao
authors.initials: C
authors.lastname: Liu
authors.affiliation: China Mobile Laboratory Response Team for Ebola in Sierra Leone, Freetown, Sierra Leone.|State Key Laboratory of Pathogen and Biosecurity, 100071, Beijing, China.|Beijing Key Laboratory of POCT for Bioemergency and Clinics, 100071, Beijing, China.
authors.forename: Ruifu
authors.initials: R
authors.lastname: Yang
authors.affiliation: Institute of Disease Control and Prevention, Academy of Military Medical Sciences, 100071, Beijing, China.|China Mobile Laboratory Response Team for Ebola in Sierra Leone, Freetown, Sierra Leone.|Key Laboratory of Zoonosis of Liaoning Province, College of Animal Science and Veterinary Medicine, Shenyang Agriculture University, 110866, Shenyang, China.
authors.forename: Zeliang
authors.initials: Z
authors.lastname: Chen
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D002648:Child; D002675:Child, Preschool; D004196:Disease Outbreaks; D029043:Ebolavirus / Q000235:genetics* / Q000302:isolation & purification; D005260:Female; D019142:Hemorrhagic Fever, Ebola / Q000175:diagnosis* / Q000453:epidemiology / Q000821:virology; D006801:Humans; D007223:Infant; D007231:Infant, Newborn; D008297:Male; D008875:Middle Aged; D021141:Nucleic Acid Amplification Techniques / Q000191:economics / Q000379:methods*; D012367:RNA, Viral / Q000235:genetics*; D012194:RNA-Directed DNA Polymerase / Q000235:genetics; D012680:Sensitivity and Specificity; D012807:Sierra Leone / Q000453:epidemiology; D013997:Time Factors
chemical_list: D012367:RNA, Viral; D012194:RNA-Directed DNA Polymerase