Repeatability Coefficients Of Variation Research Articles

Repeatability and Reproducibility of Intraocular Pressure and Dynamic Corneal Response Parameters Assessed by the Corvis ST.

Purpose To assess the repeatability and reproducibility of dynamic corneal response parameters measured by the Corvis ST (Oculus, Wetzlar, Germany). Methods One eye randomly selected from 32 healthy volunteers was examined by the Corvis ST. Three different devices were used in an alternated random order for taking three measurements at each device in each subject. Standard intraocular pressure (IOP), the biomechanical-compensated IOP (bIOP), and DCR parameters were evaluated. The within-subject standard deviation (ζw) and coefficient of variation (CV) were assessed. Results Regarding pressure indices, the ζw was below 1 mmHg for repeatability (0.98 for IOP and 0.89 for bIOP) and the CV was 6.6% for IOP and 6.1% for bIOP. For reproducibility, the ζw was around 1 mmHg (1.12 for IOP and 1.05 for bIOP) and the CV was 7.6% for IOP and 7.1% for bIOP. Most of DCR indices presented CV for repeatability below 4%. For reproducibility, the CV of most of the indices were below 6%. The deformation amplitude (DA) ratio in 1 mm and integrated radius were below 4% (1.2% and 3.8%, resp.). Conclusions The Corvis ST showed good precision (repeatability and reproducibility) for IOP measurements and for DCR in healthy eyes.

Large-scale performance evaluation of Accu-Chek inform II point-of-care glucose meters

Background: The aim of this study was to report the experience of large-scale performance evaluation of 238 Accu-Chek Inform II point-of-care (POC) glucose meters in a single medical setting.Methods: The repeatability of 238 POC devices, the within-site imprecision of 12 devices, and the linearity of 49 devices were evaluated using glucose control solutions. The glucose results of 24 POC devices and central laboratory were compared using patient samples.Results: Mean concentration of control solutions was 2.39 mmol/L for Level 1 and 16.52 mmol/L for Level 2. The pooled repeatability coefficient of variation (CV) of the 238 devices was 2.0% for Level 1 and 1.6% for Level 2. The pooled within-site imprecision CV and reproducibility CV of the 12 devices were 2.7% and 2.7% for Level 1, and 1.9%, and 1.9% for Level 2, respectively. The test results of all 49 devices were linear within analytical measurement range from 1.55–31.02 mmol/L. The correlation coefficient for individual POC devices ranged from 0.9967–0.9985. The total correlation coefficient for the 24 devices was 0.998.Conclusions: The Accu-Chek Inform II POC blood glucose meters performed well in terms of precision, linearity, and correlation evaluations. Consensus guidelines for the large-scale performance evaluations of POC devices are required.

Point-of-care PT and aPTT in patients with suspected deficiencies of coagulation factors.

There are several clinical settings and patient conditions especially in intensive care units, emergency departments, and operating theaters, where the coagulation status of a patient must be known immediately and point-of-care (POC) systems are beneficial due to low time to result. This noninterventional, single-blinded, multicenter study with prospectively collected whole blood samples was performed to evaluate the diagnostic accuracy of the CoaguChek PT Test (POC PT) and CoaguChek aPTT Test (POC aPTT) compared to standard laboratory testing in patients with suspected deficiencies of coagulation factors. In total, 390 subjects were included. Both POC PT and POC aPTT showed concordance with the laboratory PT and aPTT. Lot-to-lot variation was below 2% both for POC PT and for POC aPTT. The mean relative difference of capillary blood compared to venous blood was 0.2 % with POC PT and 8.4% with POC aPTT. The coefficients of variation for repeatability of POC PT using whole blood were found to be between 2% and 3.6%. Our findings suggest reliable quantitative results with this POC system to support on-site decision-making for patients with suspected deficiencies of coagulation factors in acute and intensive care.

The application of headspace gas chromatography coupled to tandem quadrupole mass spectrometry for the analysis of furan in baby food samples

Selective methodology employing a tandem quadrupole mass spectrometer coupled to a gas chromatograph with headspace autosampler (HS-GC–MS/MS) was elaborated in this study. Application of the elaborated procedure resulted in a limit of detection of 0.021μgkg−1 and a limit of quantification of 0.071μgkg−1. The mean recoveries during in-house validation ranged from 89% to 109%, and coefficients of variation for repeatability ranged from 4% to 11%. The proposed analytical method was applied for monitoring the furan content of 30 commercial baby food samples available on the Latvian retail market. The level of furan found in these samples varied from 0.45 to 81.9μgkg−1, indicating that infants whose sole diet comprises baby food sold in jars and cans are exposed constantly to furan. Samples containing vegetables and meat had higher levels of furan than those containing only fruits.

Spectrophotometric method for fast quantification of ascorbic acid and dehydroascorbic acid in simple matrix for kinetics measurements

A simple, rapid and reliable method was developed for quantifying ascorbic (AA) and dehydroascorbic (DHAA) acids and validated in 20mM malate buffer (pH 3.8). It consists in a spectrophotometric measurement of AA, either directly on the solution added with metaphosphoric acid or after reduction of DHAA into AA by dithiothreitol. This method was developed with real time measurement of reactions kinetics in bulk reactors in mind, and was checked in terms of linearity, limits of detection and quantification, fidelity and accuracy. The linearity was found satisfactory on the range of 0–6.95mM with limits of detection and quantification of 0.236mM and 0.467mM, respectively. The method was found acceptable in terms of fidelity and accuracy with a coefficient of variation for repeatability and reproducibility below 6% for AA and below 15% for DHAA, and with a recovery range of 97–102% for AA and 88–112% for DHAA.

Technical performance of a novel, fully automated electrochemiluminescence immunoassay for the quantitation of β-amyloid (1–42) in human cerebrospinal fluid

IntroductionAvailable assays for quantitation of the Alzheimer's disease (AD) biomarker amyloid-beta 1–42 (Aβ [1–42]) in cerebrospinal fluid demonstrate significant variability and lack of standardization to reference measurement procedures (RMPs). We report analytical performance data for the novel Elecsys β-amyloid (1–42) assay (Roche Diagnostics). MethodsLot-to-lot comparability was tested using method comparison. Performance parameters were measured according to Clinical & Laboratory Standards Institute (CLSI) guidelines. The assay was standardized to a Joint Committee for Traceability in Laboratory Medicine (JCTLM) approved RMP. ResultsLimit of quantitation was <11.28 pg/mL, and the assay was linear throughout the measuring range (200–1700 pg/mL). Excellent lot-to-lot comparability was observed (correlation coefficients [Pearson's r] >0.995; bias in medical decision area <2%). Repeatability coefficients of variation (CVs) were 1.0%–1.6%, intermediate CVs were 1.9%–4.0%, and intermodule CVs were 1.1%–3.9%. Estimated total reproducibility was 2.0%–5.1%. Correlation with the RMP was good (Pearson's r, 0.93). DiscussionThe Elecsys β-amyloid (1–42) assay has high analytical performance that may improve biomarker-based AD diagnosis.

Application of Hplc-Pda Method Using Two Different Extraction Procedures for the Determination of Alkylresorcinols in Cereals

Abstract Cereals, especially barley, are an important source of vitamins, minerals, dietary fibre and various phytochemicals, such as alkylresorcinols (ARs). Cereal ARs are a group of phenolic lipids located in the outer parts of grain, particularly in rye and wheat, but not found in refined flour or in refined products from cereals. This study focuses on the comparison of different extraction procedures applied for the determination of the content of ARs (C15:0 - C23:0) in grain of Latvian barley genotypes. The content of ARs in 1 rye and 16 barley samples grown with different amounts of fertilier was determined by High Performance Liquid Chromatography method with Photodiode Array detection (HPLC-PDA) developed by us. Two different extraction methods were compared: accelerated Soxhlet extraction and 24-hour extraction. Aside from validation of the extraction procedures, validation parameters for the HPLC-PDA based quantitation method were provided. The coefficients of variation for repeatability and intermediate precision were &lt; 9% and &lt; 3%, respectively. The content of ARs determined with the HPLC-PDA method in conjunction with accelerated Soxhlet extraction was up to 1.5 times higher than using 24-hour extraction. AR content varied from 2.11 ± 0.04 to 3.80 ± 0.10 mg·100 g-1 for 24-hour extraction and from 2.66 ± 0.06 to 5.70 ± 0.20 mg·100 g-1 for accelerated Soxhlet extraction, indicating the increased efficiency of this procedure in analysis of ARs.

Optical Coherence Tomography Angiography of Peripapillary Retinal Blood Flow Response to Hyperoxia.

To measure the change in peripapillary retinal blood flow in response to hyperoxia by using optical coherence tomography (OCT) angiography. One eye of each healthy human participants (six) was scanned with a commercial high-speed (70 kHz) spectral OCT. Scans were captured twice after 10-minute exposures to normal breathing (baseline) and hyperoxia. Blood flow was detected by the split-spectrum amplitude-decorrelation angiography (SSADA) algorithm. Peripapillary retinal blood flow index and vessel density were calculated from en face maximum projections of the retinal layers. The experiment was performed on 2 separate days for each participant. Coefficient of variation (CV) was used to measure within-day repeatability and between-day reproducibility. Paired t-tests were used to compare means of baseline and hyperoxic peripapillary retinal blood flow. A decrease of 8.87% ± 3.09% (mean ± standard deviation) in flow index and 2.61% ± 1.50% in vessel density was observed under hyperoxia. The within-day repeatability CV of baseline measurements was 5.75% for flow index and 1.67% for vessel density. The between-day reproducibility CV for baseline flow index and vessel density was 11.1% and 1.14%, respectively. The between-day reproducibility of the hyperoxic response was 3.71% and 1.67% for flow index and vessel density, respectively. Optical coherence tomography angiography with SSADA was able to detect a decrease in peripapillary retinal blood flow in response to hyperoxia. The response was larger than the variability of baseline measurements. The magnitude of an individual's hyperoxic response was highly variable between days. Thus, reliable assessment may require averaging multiple measurements.

Metabolomics study of cured tobacco using liquid chromatography with mass spectrometry: Method development and its application in investigating the chemical differences of tobacco from three growing regions

Cured tobacco is an important plant material. Component studies are a big challenge for its significantly diverse chemical properties and vastly different concentrations. In this work, liquid chromatography with quadrupole time-of-flight mass spectrometry was used to perform a metabolomics study of cured tobacco owing to its efficient separation and detection of semipolar metabolites. A solvent of methanol/water (8:2, v/v) and 30 min of ultrasound time were found to be optimal to perform extraction. 95, 92, and 93% of metabolite features had within 20% of coefficient of variation for repeatability, intraday and interday precision analysis, respectively, indicating a good stability of the method developed. 113 metabolites were identified in cured tobacco based on accurate mass, retention time, and MS/MS fragments. The developed method was applied to a metabolomics study of cured tobacco from three growing regions. Forty three metabolites were found to be contributed to the classification. It is shown that the developed method can be applied to metabolomics analysis of plant materials.

Development and evaluation of Taqman assays for the differentiation of Dickeya (sub)species

TaqMan assays were developed for the detection of seven Dickeya species, namely D. dianthicola, D. dadantii, D. paradisiaca, D. chrysanthemi, D. zeae, D. dieffenbachiae and D. solani. Sequences of the gene coding for dnaX were used for the design of primers and probes. In studies with axenic cultures of bacteria, the assays were highly specific and only reacted with strains of the target species, and not with non-target bacteria, including those belonging to other Dickeya species and other genera. The detection thresholds for DNA extracted from pure cultures of target strains ranged from 10 to 100 fg. The TaqMan assays for D. dianthicola and D. solani were more extensively evaluated as part of a method validation procedure. The threshold level for target bacteria added to a potato peel extract diluted ten-times in a semi-selective broth, was strain dependent and ranged from 1,000 to 100,000 cfu/ml. The coefficients of variation for repeatability and reproducibility were low and results were largely independent of the type of substrate, i.e. potato tuber or carnation leaf extracts. However, during routine testing of seed potatoes, false-positive reactions were found with the assay for D. solani. The use of the TaqMan assays for inspection of plant propagation material, ecological studies and studies on the effect of control strategies in disease management strategies is discussed.

Improved gas chromatography-flame ionization detector analytical method for the analysis of epoxy fatty acids

In this study an improved method for analysis of epoxy fatty acids is reported. Data obtained from analysis of polar fatty acids has previously been presented, but due to the high number of compounds that co-elute in the polar fraction, the resultant chromatograms are complex which may lead to compromising the accuracy of the data. A three steps separation of fatty acid methyl esters (FAMEs) by solid-phase extraction (SPE) on a silica gel column to remove hydroxy fatty acid interferences was proposed. This approach is opposed to a two step separation procedure that has been often used to prevent analytical interferences caused by non-altered fatty acids. A gas chromatograph with a flame ionization detector (GC-FID) equipped with a polar CP-Sil 88™ column was used. Quantification was based on the use of methyl nonadecanoate (C19:0), as an internal standard. Individual mono epoxy fatty acids were well separated without co-eluting compounds. The optimized method was finally applied to screen epoxy fatty acids in 37 fresh oil samples. Results obtained for the total epoxy fatty acids were in the range 0.03–2mgg−1 of oil with repeatability coefficient of variation (CV) ranging from 2.8 to 9.9% for duplicate analysis showing that the results obtained are repeatable.

Determination of optimal replicate number for validation of imprecision using fluorescence cell-based assays: Proposed practical method

Assay validation includes determination of inherent imprecision across the reportable range. However, specific practical guidelines for determinations of precision for cell-based fluorescence assays performed on flow cytometers are currently lacking. Replicates of 10 or 20 measurements were obtained for flow cytometric assays developed for clinical in vitro diagnostic use, including neutrophil CD64 expression for infection/sepsis detection, fetal red cell enumeration for fetomaternal hemorrhage detection, human equilibrative nucleoside transporter 1 quantitation in leukocytes for possible correlation with drug responsiveness, and CD34+ hematopoietic stem cell enumeration of apheresis products, using up to three different instrument platforms for each assay. For each assay, the mean, 95% confidence intervals (95% CIs) of the mean, standard deviation, and coefficient of variation (CV) of sequential replicates were determined. For all assays and most instrument platforms, <5 replicates were found adequate to validate assay imprecision levels below the 5-10% CV for repeatability claimed by the manufacturers of these assays. Results plotted as a novel parameter derived from the 95% CI and the cumulative mean for replicates, termed variance factor (VF), provide a data-driven means for determining optimal replicate numbers. The novel VF can provide information to guide the practical selection of optimal replicate numbers for validation of imprecision in flow cytometric assays. The optimal number of replicates was assay and instrument platform dependent. Our findings indicate that three to four replicates are sufficient for most flow cytometric assays and instrument combinations, rather than the higher numbers suggested by CLSI guidelines for soluble analytes.

Evaluation of pre-analytical and analytical factors that contribute to variability in CSF biomarker levels

development for tau that shows a marked improvement in precision, linearity and reproducibility when compared to currently available tests. Methods: An in vitro diagnostic test for tau is being developed for the VITROS Immunodiagnostic Systems, and analytical performance was evaluated following CLSI guidelines using two VITROS Tau assay reagent lots, 3 CSF pools, and 3 controls. Linearity was tested with 11 admixtures of endogenous tau in CSF and recombinant tau 441 in buffer. Interference testing included commonly prescribed drugs, recombinant tau, Ab peptides, and endogenous substances. Limit of detection (LoD) and lower limit of quantitation were confirmed with 20 replicates per day on 3 days. Results: The median within-laboratory coefficient of variation (CV) for the 3 CSF pools was 3.3% with a range of 2.4% 4.2%. The controls’ within-laboratory CV was 1.2%. Repeatability CVs were 1.9% for CSF pools and 1.2% for controls. The assay showed good linearity across themeasuring range (08,000pg/mL) with the admixture of endogenous and synthetic peptide. Bias introduced by commonly prescribed drugs at high levels was <610%. The interference from Ab peptides at greater than known physiological concentrations as well as from endogenous substances was <5%. The limit of detection was 9.5pg/mL; the lower limit of quantitation was 40pg/mL. Conclusions: VITROS Immunodiagnostic Products Tau Assay 1 demonstrates excellent analytical performance: it is precise, linear and the tested interferences cause no significant bias. The fully automated format and ability to perform these assays in a clinical laboratory should make testing for these important markers more reliable and routine.

Determination of optimal replicate number for validation of imprecision using fluorescence cell based assays: Proposed practical method.

Background: Assay validation includes determination of inherent imprecision across the reportable range. However specific practical guidelines for determinations of precision for cell based fluorescence assays performed on flow cytometers are currently lacking. Methods: Replicates of 10 or 20 measurements were obtained for flow cytometric assays developed for clinical IVD use, including neutrophil CD64 expression for infection/sepsis detection, fetal red cell enumeration for fetomaternal hemorrhage detection, human equilibrative nucleoside transporter 1 (hENT1) quantitation in leukocytes for possible correlation with drug responsiveness, and CD34+ hematopoietic stem cell (HSC) enumeration of apheresis products, using up to three different instrument platforms for each assay. For each assay, the mean, 95% confidence intervals of the mean (95%CI), standard deviation and coefficient of variation (CV) of sequential replicates were determined. Results: For all assays and most instrument platforms <5 replicates were found adequate to validate assay imprecision levels below the 5-10% CV for repeatability claimed by the manufacturers of these assays. Results plotted as a novel parameter derived from the 95%CI and the cumulative mean for replicates, termed variance factor (VF), provide a data driven means for determining optimal replicate numbers. Conclusions: The novel VF can provide information to guide the practical selection of optimal replicate numbers for validation of imprecision in flow cytometric assays. The optimal number of replicates was assay and instrument platform dependent. Our findings indicate 3-4 replicates are sufficient for most flow cytometric assays and instrument combinations, rather than the higher numbers suggested by CLSI guidelines for soluble analytes. © 2013 Clinical Cytometry Society.

Determination of macrocyclic lactones in food and feed

A confirmatory high-performance liquid chromatography method with fluorescence detection (HPLC-FLD) was developed and validated for the simultaneous determination of the following anti-parasitic veterinary drugs in foodstuffs (liver, muscle and milk) and feed: abamectin, doramectin, emamectin, eprinomectin, ivermectin and moxidectin. Samples were extracted, purified and analysed by HPLC-FLD after prior derivatisation of analytes with N-methylimidazole, trifluoracetic anhydride and acetic acid to produce stable fluorescent derivatives. Recoveries were in the range of 73–98% (liver), 82–93% (muscle), 77–84% (milk), and 93–110% (feed). In all matrices the coefficients of variation for repeatability and intra-laboratory reproducibility were satisfactory. For emamectin in liver the worst method performance was observed. As this multi-analyte method fully met the required criteria and applicability both to food and feed, it has been successfully applied in National Control Plans in food and feed.

Reproducibility of the anti-Factor Xa and anti-Factor IIa assays applied to enoxaparin solution

Enoxaparin is a widely used subcutaneously administered antithrombotic agent comprising a complex mixture of glycosaminoglycan chains. Owing to this complexity, its antithrombotic potency cannot be defined by physicochemical methods and is therefore evaluated using an enzymatic assay of anti-Xa and anti-IIa activity. Maintaining consistent anti-Xa activity in the final medicinal product allows physicians to ensure administration of the appropriate dosage to their patients. Bioassays are usually complex and display poorer reproducibility than physicochemical tests such as HPLC assays. Here, we describe the implementation of a common robotic platform and standard release potency testing procedures for enoxaparin sodium injection (Lovenox®, Sanofi, Paris, France) products at seven quality control sites within Sanofi. Qualification and analytical procedures, as well as data handling, were optimized and harmonized to improve assay reproducibility. An inter-laboratory study was performed in routine-release conditions. The coefficients of variation for repeatability and reproducibility in assessments of anti-Xa activity were 1.0% and 1.2%, respectively. The tolerance interval in reproducibility precision conditions, expressed as percentage potency, was 96.8–103.2% of the drug product target of 10,000IU/ml, comparing favorably with the United States of America Pharmacopeia specification (90–110%). The maximum difference between assays in two different laboratories is expected to be 4.1%. The reproducibility characteristics of anti-IIa activity assessments were found to be similar. These results demonstrate the effectiveness of the standardization process established and allow for further improvements to quality control in Lovenox® manufacture. This process guarantees closeness between actual and target potencies, as exemplified by the results of release assays obtained during a three-year period.

Performance characteristics of a rapid method for iodine determination in milk / Szybka metoda oznaczania jodu w mleku – walidacja i akredytacja zgodnie z normą ISO/IEC 17025:2005

Streszczenie Badania powtarzalności i odtwarzalności przeprowadzono na 72 próbkach. Określono najważniejsze parametry walidacji, w tym składowe niepewności. Wyniki analiz porównano z wartościami referencyjnymi. Powtarzalność i odtwarzalność metody nie przekraczały 8 i 10%, odpowiednio. LOQ w oznaczanym roztworze próbki mleka wynosił 14 μg/L. Zakres roboczy krzywej kalibracji wynosił od 7 do 200 μg/L. Niepewność metody (P≤0,05) uwzględniająca błędy odtwarzalności wewnątrzlaboratoryjnej, odzysku, czystości wzorca oraz błędy szkła miarowego wynosiła (%) 17.8 i 22.8 (n = 2 i n = 1). Podczas wykonywania rutynowych analiz powinien być sprawdzany współczynnik zmienności dla powtarzalności, który nie powinien przekraczać granicy powtarzalności wynoszącej 16%. Metoda cechuje się wystarczającą wiarygodnością, co zostało potwierdzone wynikami walidacji. Procedura jest szybka, prosta i ma niską wartość LOQ. Metodę akredytowano zgodnie z normą ISO/IEC 17025:2005.

Inter-laboratory trials for analysis of perfluorooctanesulfonate and perfluorooctanoate in water samples: Performance and recommendations

The ISO 25101 (International Organization for Standardization, Geneva) describes a new international standard method for the determination of perfluorooctanesulfonate (PFOS) and perfluorooctanoate (PFOA) in unfiltered samples of drinking and surface waters. The method is based on the extraction of target analytes by solid phase extraction, solvent elution, and determination by high performance liquid chromatography–tandem mass spectrometry (HPLC–MS/MS). For the determination of the performance of this method, more than 20 laboratories from 9 different countries participated in an inter-laboratory trial in 2006. In addition, inter-laboratory trials were conducted in 2008 and 2009 for the analysis of perfluoroalkylsubstances (PFASs), including PFOS and PFOA, in water samples by following the protocols of Japanese Industrial Standard (JIS). Overall, the repeatability coefficients of variation (i.e., within-laboratory precision) for PFOS and PFOA in all water samples were between 3 and 11%, showing a adequate precision of the ISO and JIS methods. The reproducibility coefficients of variation (i.e., between-laboratory precision) were found to vary within a range of 7–31% for surface water and 20–40% for wastewater. The recoveries of PFOS and PFOA, as a measure of accuracy, varied from 84 to 100% for surface water and from 84 to 100% for wastewater among the samples with acceptable criteria for internal standards recovery. The determined concentrations of PFASs in samples compared well with the “true” values. The results of the inter-laboratory trial confirmed that the analytical methods are robust and reliable and can be used as a standard method for the analysis of target compounds in water samples.

Validation of a Method for Determining Cholesterol in Egg Yolks/ Walidacja metody oznaczania cholesterolu w żółtkach jaj

Abstract The aim of the study was to validate a gas chromatographic method for determining cholesterol in egg yolks according to the EN ISO/IEC 17025 standard. Of the two methods, with and without internal standard, the former was characterized by lower uncertainty, with a repeatability of 4% and within-laboratory reproducibility of 6%. The method’s uncertainty (n = 2, P≤0.05), which included sample preparation errors and chromatographic measurement errors, was 10.6%. Mean recovery was 99.9% and limit of quantification was 0.16 mg/g. The coefficient of variation for repeatability, which is calculated during routine analyses, should not exceed the 8% limit of repeatability. The method is reliable, as confirmed by the results of validation, and the procedure is relatively rapid and simple.

Validation of an enzyme-linked immunosorbent assay for the quantification of human IgG directed against the repeat region of the circumsporozoite protein of the parasite Plasmodium falciparum

BackgroundSeveral pre-erythrocytic malaria vaccines based on the circumsporozoite protein (CSP) antigen of Plasmodium falciparum are in clinical development. Vaccine immunogenicity is commonly evaluated by the determination of anti-CSP antibody levels using IgG-based assays, but no standard assay is available to allow comparison of the different vaccines.MethodsThe validation of an anti-CSP repeat region enzyme-linked immunosorbent assay (ELISA) is described. This assay is based on the binding of serum antibodies to R32LR, a recombinant protein composed of the repeat region of P. falciparum CSP. In addition to the original recombinant R32LR, an easy to purify recombinant His-tagged R32LR protein has been constructed to be used as solid phase antigen in the assay. Also, hybridoma cell lines have been generated producing human anti-R32LR monoclonal antibodies to be used as a potential inexhaustible source of anti-CSP repeats standard, instead of a reference serum.ResultsThe anti-CSP repeats ELISA was shown to be robust, specific and linear within the analytical range, and adequately fulfilled all validation criteria as defined in the ICH guidelines. Furthermore, the coefficient of variation for repeatability and intermediate precision did not exceed 23%. Non-interference was demonstrated for R32LR-binding sera, and the assay was shown to be stable over time.ConclusionsThis ELISA, specific for antibodies directed against the CSP repeat region, can be used as a standard assay for the determination of humoral immunogenicity in the development of any CSP-based P. falciparum malaria vaccine.