Recombinase polymerase amplification and lateral flow assay for ultrasensitive detection of low-density Plasmodium falciparum infection from controlled human malaria infection studies and naturally acquired infections
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Recombinase polymerase amplification and lateral flow assay for ultrasensitive detection of low-density Plasmodium falciparum infection from controlled human malaria infection studies and naturally acquired infections. / Lalremruata, Albert; Nguyen, The Trong; McCall, Matthew B.B.; Mombo-Ngoma, Ghyslain; Agnandji, Selidji T.; Adegnika, Ayôla A.; Lell, Bertrand; Ramharter, Michael; Hoffman, Stephen L.; Kremsner, Peter G.; Mordmüller, Benjamin.
in: Journal of Clinical Microbiology, Jahrgang 58, Nr. 5, e01879-19, 23.04.2020.Publikationen: SCORING: Beitrag in Fachzeitschrift/Zeitung › SCORING: Zeitschriftenaufsatz › Forschung › Begutachtung
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TY - JOUR
T1 - Recombinase polymerase amplification and lateral flow assay for ultrasensitive detection of low-density Plasmodium falciparum infection from controlled human malaria infection studies and naturally acquired infections
AU - Lalremruata, Albert
AU - Nguyen, The Trong
AU - McCall, Matthew B.B.
AU - Mombo-Ngoma, Ghyslain
AU - Agnandji, Selidji T.
AU - Adegnika, Ayôla A.
AU - Lell, Bertrand
AU - Ramharter, Michael
AU - Hoffman, Stephen L.
AU - Kremsner, Peter G.
AU - Mordmüller, Benjamin
N1 - Copyright © 2020 American Society for Microbiology.
PY - 2020/4/23
Y1 - 2020/4/23
N2 - Microscopy and rapid diagnostic tests (RDTs) are the main diagnostic tools for malaria but fail to detect low-density parasitemias that are important for maintaining malaria transmission. To complement existing diagnostic methods, an isothermal reverse transcription-recombinase polymerase amplification and lateral flow assay (RT-RPA) was developed. We compared the performance with that of ultrasensitive reverse transcription-quantitative PCR (uRT-qPCR) using nucleic acid extracts from blood samples (n=114) obtained after standardized controlled human malaria infection (CHMI) with Plasmodium falciparum sporozoites. As a preliminary investigation, we also sampled asymptomatic individuals (n=28) in an area of malaria endemicity (Lambaréné, Gabon) to validate RT-RPA and assess its performance with unprocessed blood samples (dbRT-RPA). In 114 samples analyzed from CHMI trials, the positive percent agreement to uRT-qPCR was 90% (95% confidence interval [CI], 80 to 96). The negative percent agreement was 100% (95% CI, 92 to 100). The lower limit of detection was 64 parasites/ml. In Gabon, RT-RPA was 100% accurate with asymptomatic volunteers (n=28), while simplified dbRT-RPA showed 89% accuracy. In a subgroup analysis, RT-RPA detected 9/10 RT-qPCR-positive samples, while loop-mediated isothermal amplification (LAMP) detected 2/10. RT-RPA is a reliable diagnostic test for asymptomatic low-density infections. It is particularly useful in settings where uRT-qPCR is difficult to implement.
AB - Microscopy and rapid diagnostic tests (RDTs) are the main diagnostic tools for malaria but fail to detect low-density parasitemias that are important for maintaining malaria transmission. To complement existing diagnostic methods, an isothermal reverse transcription-recombinase polymerase amplification and lateral flow assay (RT-RPA) was developed. We compared the performance with that of ultrasensitive reverse transcription-quantitative PCR (uRT-qPCR) using nucleic acid extracts from blood samples (n=114) obtained after standardized controlled human malaria infection (CHMI) with Plasmodium falciparum sporozoites. As a preliminary investigation, we also sampled asymptomatic individuals (n=28) in an area of malaria endemicity (Lambaréné, Gabon) to validate RT-RPA and assess its performance with unprocessed blood samples (dbRT-RPA). In 114 samples analyzed from CHMI trials, the positive percent agreement to uRT-qPCR was 90% (95% confidence interval [CI], 80 to 96). The negative percent agreement was 100% (95% CI, 92 to 100). The lower limit of detection was 64 parasites/ml. In Gabon, RT-RPA was 100% accurate with asymptomatic volunteers (n=28), while simplified dbRT-RPA showed 89% accuracy. In a subgroup analysis, RT-RPA detected 9/10 RT-qPCR-positive samples, while loop-mediated isothermal amplification (LAMP) detected 2/10. RT-RPA is a reliable diagnostic test for asymptomatic low-density infections. It is particularly useful in settings where uRT-qPCR is difficult to implement.
KW - CHMI
KW - Diagnostics
KW - Elimination
KW - Isothermal
KW - Mass screen and treat
KW - Plasmodium falciparum
KW - Submicroscopic
KW - Malaria, Falciparum/diagnosis
KW - Plasmodium falciparum/genetics
KW - Humans
KW - Recombinases
KW - Malaria/diagnosis
KW - Nucleic Acid Amplification Techniques
KW - Gabon
KW - Molecular Diagnostic Techniques
KW - Sensitivity and Specificity
UR - http://www.scopus.com/inward/record.url?scp=85084027252&partnerID=8YFLogxK
U2 - 10.1128/JCM.01879-19
DO - 10.1128/JCM.01879-19
M3 - SCORING: Journal article
C2 - 32102854
AN - SCOPUS:85084027252
VL - 58
JO - J CLIN MICROBIOL
JF - J CLIN MICROBIOL
SN - 0095-1137
IS - 5
M1 - e01879-19
ER -