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25OHD analogues and vacuum blood collection tubes dramatically affect the accuracy of automated immunoassays.

Yu S, Cheng X, Fang H, Zhang R, Han J, Qin X, Cheng Q, Su W, Hou L, Xia L, Qiu L - Sci Rep (2015)

Bottom Line: However, the Siemens system produced a large positive mean bias of 12.5 ng/mL and a poor Kappa value when using tubes with clot activator and gel separator.Serum 25OHD in VACUETTE tubes with gel and clot activator, as measured by the Siemens system, produced significantly higher values than did samples collected in VACUETTE tubes with no additives.Bias decreased and clinical agreement improved significantly when using tubes with no additives.

View Article: PubMed Central - PubMed

Affiliation: Department of Clinical Laboratory, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences, Beijing 100730, China.

ABSTRACT
Variations in vitamin D quantification methods are large, and influences of vitamin D analogues and blood collection methods have not been systematically examined. We evaluated the effects of vitamin D analogues 25OHD2 and 3-epi 25OHD3 and blood collection methods on vitamin D measurement, using five immunoassay systems and liquid chromatography-tandem mass spectrometry (LC-MS/MS). Serum samples (332) were selected from routine vitamin D assay requests, including samples with or without 25OHD2 or 3-epi 25OHD3, and analysed using various immunoassay systems. In samples with no 25OHD2 or 3-epi 25OHD3, all immunoassays correlated well with LC-MS/MS. However, the Siemens system produced a large positive mean bias of 12.5 ng/mL and a poor Kappa value when using tubes with clot activator and gel separator. When 25OHD2 or 3-epi 25OHD3 was present, correlations and clinical agreement decreased for all immunoassays. Serum 25OHD in VACUETTE tubes with gel and clot activator, as measured by the Siemens system, produced significantly higher values than did samples collected in VACUETTE tubes with no additives. Bias decreased and clinical agreement improved significantly when using tubes with no additives. In conclusion, most automated immunoassays showed acceptable correlation and agreement with LC-MS/MS; however, 25OHD analogues and blood collection tubes dramatically affected accuracy.

No MeSH data available.


Related in: MedlinePlus

Comparison of DiaSorin and LC-MS/MS.A–E are Passing-Bablok regression analyses for 166 serum samples containing only 25OHD3, 111 serum samples containing both 25OHD3 and 25OHD2, 31 serum samples containing 25OHD3, and all 332 serum samples, respectively. F–J are Bland-Altman plots showing the bias between DiaSorin and LC-MS/MS for 166 serum samples containing only 25OHD3; 111 serum samples containing both 25OHD3 and 25OHD2; 31 serum samples containing 25OHD3; 24 samples containing 25OHD3, 25OHD2, and 3-epi 25OHD3; and all 332 serum samples, respectively.
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f2: Comparison of DiaSorin and LC-MS/MS.A–E are Passing-Bablok regression analyses for 166 serum samples containing only 25OHD3, 111 serum samples containing both 25OHD3 and 25OHD2, 31 serum samples containing 25OHD3, and all 332 serum samples, respectively. F–J are Bland-Altman plots showing the bias between DiaSorin and LC-MS/MS for 166 serum samples containing only 25OHD3; 111 serum samples containing both 25OHD3 and 25OHD2; 31 serum samples containing 25OHD3; 24 samples containing 25OHD3, 25OHD2, and 3-epi 25OHD3; and all 332 serum samples, respectively.

Mentions: Although the DiaSorin system was supposed to detect 25OHD with 100% efficiency and specificity for 25OHD3, the correlation coefficient declined, the bias increased, and the Kappa value decreased significantly when the samples contained 25OHD2 or 3-epi 25OHD3 (Fig. 2). The other four immunoassays exhibited a similar trend (Table 2). The level of 25OHD2 correlated significantly with the bias between immunoassay methods (for Abbott, DiaSorin, IDS, Roche, and Siemens: r = 0.848, 0.909, 0.834, 0.849, and 0.282, respectively) and LC-MS/MS (Supplemental Figure 2). And it was shown that when there was no 25OHD2, bias% at the medical decision level of Roche was the smallest, however, when 25OHD2 present, bias% at medical decision level significantly increased(Supplemental Table 2).


25OHD analogues and vacuum blood collection tubes dramatically affect the accuracy of automated immunoassays.

Yu S, Cheng X, Fang H, Zhang R, Han J, Qin X, Cheng Q, Su W, Hou L, Xia L, Qiu L - Sci Rep (2015)

Comparison of DiaSorin and LC-MS/MS.A–E are Passing-Bablok regression analyses for 166 serum samples containing only 25OHD3, 111 serum samples containing both 25OHD3 and 25OHD2, 31 serum samples containing 25OHD3, and all 332 serum samples, respectively. F–J are Bland-Altman plots showing the bias between DiaSorin and LC-MS/MS for 166 serum samples containing only 25OHD3; 111 serum samples containing both 25OHD3 and 25OHD2; 31 serum samples containing 25OHD3; 24 samples containing 25OHD3, 25OHD2, and 3-epi 25OHD3; and all 332 serum samples, respectively.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4588576&req=5

f2: Comparison of DiaSorin and LC-MS/MS.A–E are Passing-Bablok regression analyses for 166 serum samples containing only 25OHD3, 111 serum samples containing both 25OHD3 and 25OHD2, 31 serum samples containing 25OHD3, and all 332 serum samples, respectively. F–J are Bland-Altman plots showing the bias between DiaSorin and LC-MS/MS for 166 serum samples containing only 25OHD3; 111 serum samples containing both 25OHD3 and 25OHD2; 31 serum samples containing 25OHD3; 24 samples containing 25OHD3, 25OHD2, and 3-epi 25OHD3; and all 332 serum samples, respectively.
Mentions: Although the DiaSorin system was supposed to detect 25OHD with 100% efficiency and specificity for 25OHD3, the correlation coefficient declined, the bias increased, and the Kappa value decreased significantly when the samples contained 25OHD2 or 3-epi 25OHD3 (Fig. 2). The other four immunoassays exhibited a similar trend (Table 2). The level of 25OHD2 correlated significantly with the bias between immunoassay methods (for Abbott, DiaSorin, IDS, Roche, and Siemens: r = 0.848, 0.909, 0.834, 0.849, and 0.282, respectively) and LC-MS/MS (Supplemental Figure 2). And it was shown that when there was no 25OHD2, bias% at the medical decision level of Roche was the smallest, however, when 25OHD2 present, bias% at medical decision level significantly increased(Supplemental Table 2).

Bottom Line: However, the Siemens system produced a large positive mean bias of 12.5 ng/mL and a poor Kappa value when using tubes with clot activator and gel separator.Serum 25OHD in VACUETTE tubes with gel and clot activator, as measured by the Siemens system, produced significantly higher values than did samples collected in VACUETTE tubes with no additives.Bias decreased and clinical agreement improved significantly when using tubes with no additives.

View Article: PubMed Central - PubMed

Affiliation: Department of Clinical Laboratory, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences, Beijing 100730, China.

ABSTRACT
Variations in vitamin D quantification methods are large, and influences of vitamin D analogues and blood collection methods have not been systematically examined. We evaluated the effects of vitamin D analogues 25OHD2 and 3-epi 25OHD3 and blood collection methods on vitamin D measurement, using five immunoassay systems and liquid chromatography-tandem mass spectrometry (LC-MS/MS). Serum samples (332) were selected from routine vitamin D assay requests, including samples with or without 25OHD2 or 3-epi 25OHD3, and analysed using various immunoassay systems. In samples with no 25OHD2 or 3-epi 25OHD3, all immunoassays correlated well with LC-MS/MS. However, the Siemens system produced a large positive mean bias of 12.5 ng/mL and a poor Kappa value when using tubes with clot activator and gel separator. When 25OHD2 or 3-epi 25OHD3 was present, correlations and clinical agreement decreased for all immunoassays. Serum 25OHD in VACUETTE tubes with gel and clot activator, as measured by the Siemens system, produced significantly higher values than did samples collected in VACUETTE tubes with no additives. Bias decreased and clinical agreement improved significantly when using tubes with no additives. In conclusion, most automated immunoassays showed acceptable correlation and agreement with LC-MS/MS; however, 25OHD analogues and blood collection tubes dramatically affected accuracy.

No MeSH data available.


Related in: MedlinePlus