A ring trial to harmonize Toxoplasma gondii microsatellite typing: comparative analysis of results and recommendations for optimization.
A ring trial to harmonize Toxoplasma gondii microsatellite typing: comparative analysis of results and recommendations for optimization.
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DOI：
10.1007/s10096-023-04597-7
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发表时间：
2023-07
影响因子：
4.5
通讯作者：
Schares, G.
中科院分区：
文献类型：
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作者：
Joeres, M.;Cardron, G.;Passebosc-Faure, K.;Plault, N.;Fernandez-Escobar, M.;Hamilton, C. M.;O'Brien-Anderson, L.;Calero-Bernal, R.;Galal, L.;Luttermann, C.;Maksimov, P.;Conraths, F. J.;Darde, M. L.;Ortega-Mora, L. M.;Jokelainen, P.;Mercier, A.;Schares, G.
为了在人畜共患寄生虫弓形虫的微卫星（MS）分型方面达到一致性，组织了五个欧洲实验室之间的环形试验。每个实验室按照先前发表的基于15个MS标记的片段长度多态性的方法循环和分析三个样品集。第一个样本集比较了总体的分型结果，并着重于DNA浓度的影响;第二个样本集着重于可以区分T.同一原型谱系内的弓形虫株;第三组集中于非原型基因型。实验室之间的方法学差异，包括用于确定MS片段长度的软件程序，使用问卷进行整理。总体而言，谱系水平分型结果达到了高度一致，特别是在DNA浓度最高的样本中。然而，对于特定的标记物，观察到实验室特定的差异。片段长度的主要中值差异（最多6个碱基对）与用于标记片段特异性引物的荧光团有关。此外，从不同供应商获得的具有相同序列的引物对产生不同长度的片段。此外，评估和解释测序图谱的方式差异可能导致片段长度测定结果偏离。例如，通过使用相同的荧光团或通过对确定的片段长度进行数值调整来协调MS分型，可以提高实验室之间结果的一致性。这是第一次实验室间比较，为优化该技术提供了指南（作为补充）。在线版本包含补充材料，可通过10.1007/s10096-023-04597-7获得。
A ring trial among five European laboratories was organized to reach consistency in microsatellite (MS) typing of the zoonotic parasite Toxoplasma gondii. Three sample sets were circulated and analyzed by each laboratory following a previously published method that is based on fragment length polymorphism of 15 MS markers. The first sample set compared typing results in general and focused on effects of DNA concentration; the second sample set focused on the polymorphic fingerprinting markers that can differentiate T. gondii strains within the same archetypal lineage; and the third set focused on non-archetypal genotypes. Methodological variations between laboratories, including the software programs used to determine MS fragment length, were collated using a questionnaire. Overall, lineage-level typing results reached a high level of agreement, especially in samples with the highest DNA concentrations. However, laboratory-specific differences were observed for particular markers. Major median differences in fragment length, of up to 6 base pairs, were related to the fluorophore used to label fragment-specific primers. In addition, primer pairs with identical sequences obtained from different suppliers resulted in fragments of differing length. Furthermore, differences in the way the sequencing profiles were assessed and interpreted may have led to deviating results in fragment length determination. Harmonization of MS typing, for example, by using the same fluorophores or by numerical adjustments applied to the fragment-lengths determined, could improve the uniformity of the results across laboratories. This is the first interlaboratory comparison, providing guidelines (added as a supplement) for the optimization of this technique. The online version contains supplementary material available at 10.1007/s10096-023-04597-7.
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影响因子：
3.8
作者：
Khan A;Ajzenberg D;Mercier A;Demar M;Simon S;Dardé ML;Wang Q;Verma SK;Rosenthal BM;Dubey JP;Sibley LD
通讯作者：
Sibley LD
影响因子：
3.8
作者：
Hamidović A;Etougbétché JR;Tonouhewa ABN;Galal L;Dobigny G;Houémènou G;Da Zoclanclounon H;Amagbégnon R;Laleye A;Fievet N;Piry S;Berthier K;Pena HFJ;Dardé ML;Mercier A
通讯作者：
Mercier A
影响因子：
7.7
作者：
Lilue J;Müller UB;Steinfeldt T;Howard JC
通讯作者：
Howard JC
影响因子：
19
作者：
Bouwknegt, Martijn;Devleesschauwer, Brecht;van der Giessen, Joke W. B.
通讯作者：
van der Giessen, Joke W. B.
影响因子：
16.6
作者：
通讯作者：
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