Analytical validation of a real-time hydrolysis probe PCR assay for quantifying Plasmodium falciparum parasites in experimentally infected human adults.
Hydrolysis probe
MIQE
Plasmodium falciparum
Quantitative PCR
Validation
Volunteer infection studies
Journal
Malaria journal
ISSN: 1475-2875
Titre abrégé: Malar J
Pays: England
ID NLM: 101139802
Informations de publication
Date de publication:
10 Apr 2021
10 Apr 2021
Historique:
received:
14
04
2020
accepted:
29
03
2021
entrez:
11
4
2021
pubmed:
12
4
2021
medline:
9
9
2021
Statut:
epublish
Résumé
Volunteer infection studies have become a standard model for evaluating drug efficacy against Plasmodium infections. Molecular techniques such as qPCR are used in these studies due to their ability to provide robust and accurate estimates of parasitaemia at increased sensitivity compared to microscopy. The validity and reliability of assays need to be ensured when used to evaluate the efficacy of candidate drugs in clinical trials. A previously described 18S rRNA gene qPCR assay for quantifying Plasmodium falciparum in blood samples was evaluated. Assay performance characteristics including analytical sensitivity, reportable range, precision, accuracy and specificity were assessed using experimental data and data compiled from phase 1 volunteer infection studies conducted between 2013 and 2019. Guidelines for validation of laboratory-developed molecular assays were followed. The reportable range was 1.50 to 6.50 log The 18S rRNA gene qPCR assay is specific and highly reproducible and can provide reliable and accurate parasite quantification. The assay is considered fit for use in evaluating drug efficacy in malaria clinical trials.
Sections du résumé
BACKGROUND
BACKGROUND
Volunteer infection studies have become a standard model for evaluating drug efficacy against Plasmodium infections. Molecular techniques such as qPCR are used in these studies due to their ability to provide robust and accurate estimates of parasitaemia at increased sensitivity compared to microscopy. The validity and reliability of assays need to be ensured when used to evaluate the efficacy of candidate drugs in clinical trials.
METHODS
METHODS
A previously described 18S rRNA gene qPCR assay for quantifying Plasmodium falciparum in blood samples was evaluated. Assay performance characteristics including analytical sensitivity, reportable range, precision, accuracy and specificity were assessed using experimental data and data compiled from phase 1 volunteer infection studies conducted between 2013 and 2019. Guidelines for validation of laboratory-developed molecular assays were followed.
RESULTS
RESULTS
The reportable range was 1.50 to 6.50 log
CONCLUSIONS
CONCLUSIONS
The 18S rRNA gene qPCR assay is specific and highly reproducible and can provide reliable and accurate parasite quantification. The assay is considered fit for use in evaluating drug efficacy in malaria clinical trials.
Identifiants
pubmed: 33838672
doi: 10.1186/s12936-021-03717-y
pii: 10.1186/s12936-021-03717-y
pmc: PMC8035755
doi:
Substances chimiques
RNA, Protozoan
0
RNA, Ribosomal, 18S
0
Types de publication
Journal Article
Validation Study
Langues
eng
Sous-ensembles de citation
IM
Pagination
181Subventions
Organisme : Medicines for Malaria Venture
ID : OPP1111147
Références
Nat Rev Immunol. 2011 Jan;11(1):57-64
pubmed: 21179119
Antimicrob Agents Chemother. 2015 Jul;59(7):4249-59
pubmed: 25963983
Trop Med Health. 2019 Aug 27;47:49
pubmed: 31485189
Mol Biochem Parasitol. 2009 Feb;163(2):123-6
pubmed: 19027798
Am J Trop Med Hyg. 2019 Jun;100(6):1466-1476
pubmed: 31017084
Sci Rep. 2016 Dec 16;6:39183
pubmed: 27982132
Vaccine. 2017 Dec 18;35(51):7065-7069
pubmed: 29153778
Antimicrob Agents Chemother. 2013 Dec;57(12):5792-807
pubmed: 24002099
PLoS One. 2014 May 16;9(5):e97398
pubmed: 24838112
J Appl Microbiol. 2012 Nov;113(5):1014-26
pubmed: 22747964
Annu Rev Pharmacol Toxicol. 2018 Jan 6;58:567-582
pubmed: 28992431
Malar J. 2011 Feb 28;10:48
pubmed: 21352599
Lancet Infect Dis. 2016 Mar;16(3):357-65
pubmed: 26774243
Korean J Parasitol. 2009 Jun;47(2):93-102
pubmed: 19488414
Clin Chem. 2009 Apr;55(4):611-22
pubmed: 19246619
Am J Trop Med Hyg. 2004 Aug;71(2):196-201
pubmed: 15306710
Clin Microbiol Rev. 2010 Jul;23(3):550-76
pubmed: 20610823
J Clin Microbiol. 2014 Sep;52(9):3303-9
pubmed: 24989601
Trends Parasitol. 2016 Mar;32(3):187-196
pubmed: 26826784
Malar J. 2016 Aug 05;15(1):398
pubmed: 27495296
J Infect Dis. 2018 Feb 14;217(5):693-702
pubmed: 29216395
Am J Trop Med Hyg. 2017 Jul;97(1):188-198
pubmed: 28719294
Am J Trop Med Hyg. 2013 Nov;89(5):824-39
pubmed: 24062484
PLoS One. 2011;6(8):e21914
pubmed: 21887214
Biomol Detect Quantif. 2017 Apr 29;12:1-6
pubmed: 28702366
J Infect Dis. 2020 Mar 2;221(6):963-972
pubmed: 31679015
Am J Trop Med Hyg. 2012 Mar;86(3):383-94
pubmed: 22403305
Antimicrob Agents Chemother. 2019 Dec 20;64(1):
pubmed: 31685476
Appl Environ Microbiol. 1997 Oct;63(10):3741-51
pubmed: 9327537
Clin Chem. 1981 Mar;27(3):493-501
pubmed: 7471403
Malar J. 2018 Jul 5;17(1):249
pubmed: 29976199
J Antimicrob Chemother. 2016 Sep;71(9):2620-7
pubmed: 27272721
Antimicrob Agents Chemother. 2017 Jan 24;61(2):
pubmed: 27872070
Malar J. 2015 Dec 23;14:520
pubmed: 26701778
Malar J. 2014 Jul 28;13:288
pubmed: 25066459
J Clin Invest. 2018 Apr 2;128(4):1551-1562
pubmed: 29389671
Malar J. 2015 Mar 18;14:117
pubmed: 25889522
J Clin Invest. 2020 Jun 1;130(6):2920-2927
pubmed: 32045385
Malar J. 2006 Dec 14;5:121
pubmed: 17169142
N Engl J Med. 2014 Jul 31;371(5):411-23
pubmed: 25075834
Lancet Infect Dis. 2018 Aug;18(8):874-883
pubmed: 29909069
Croat Med J. 2009 Jun;50(3):207-17
pubmed: 19480017
Methods Mol Biol. 2012;820:7-23
pubmed: 22131023
Sex Transm Infect. 2009 Apr;85(2):102-5
pubmed: 19004866
Nat Rev Microbiol. 2014 Dec;12(12):833-40
pubmed: 25329408
J Clin Microbiol. 2001 Feb;39(2):485-93
pubmed: 11158094
PLoS One. 2014 Nov 18;9(11):e109654
pubmed: 25405724