Evaluation of HBs Ag, HCV and HIV Ag-Ab Assays using Bio-Rad Elite Microplate Analyzer

Sang-Hyun Hwang; Heung-Bum Oh; Hyon-Suk Kim; Eun Yup Lee

doi:10.3343/kjlm.2006.26.6.436

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Hwang, Oh, Kim, and Lee: Evaluation of HBs Ag, HCV and HIV Ag-Ab Assays using Bio-Rad Elite Microplate Analyzer

Original Article

Korean J Leg Med 2006;26(6):436-441.

Published online: 10 February 2006

DOI: https://doi.org/10.3343/kjlm.2006.26.6.436

Evaluation of HBs Ag, HCV and HIV Ag-Ab Assays using Bio-Rad Elite Microplate Analyzer

Sang-Hyun Hwang, M.D.¹, Heung-Bum Oh, M.D.², Hyon-Suk Kim, M.D.³, Eun Yup Lee, M.D.¹

¹Department of Laboratory Medicine, Pusan National University School of Medicine, Busan, Korea

²Department of Laboratory Medicine, University of Ulsan College of Medicine and Asan Medical Center, Seoul, Korea

³Department of Laboratory Medicine, Yonsei University College of Medicine, Seoul, Korea

Received 7 August 2006 Revised 25 October 2006 Accepted 12 November 2006

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

Background

In this study, we evaluated the performance of Elite microplate analyzer (Bio-Rad Laboratories, France) and the related assays (ULTRA line) for the detection of hepatitis B virus (HBV), hepatitis C virus (HCV) and human immunodeficiency virus (HIV).

Methods

Seroconversion panels, HBsAg positive/HBsAg negative (n=99/n=1,422), anti-HCV positive/negative (n=97/n=1,670), and anti-HIV positive/negative (n=112/n=1,704) samples were used to evaluate the performance of Monolisa HBsAg ULTRA, Monolisa HCV Ag-Ab ULTRA, and Genscreen ULTRA HIV Ag-Ab, respectively. The agreement of Elite microplate analyzer with CODA analyzer (Bio-Rad Laboratories, CA, USA) was also evaluated.

Results

The detection limit of Monolisa HBsAg ULTRA was 0.034 IU/mL. For Monolisa HBsAg ULTRA, Monolisa HCV Ag-Ab ULTRA, and Genscreen ULTRA HIV Ag-Ab, diagnostic sensitivities were all 100%, diagnostic specificities were 100%, 99.8% and 99.9%, and total CVs (coefficients of variation) were 13.8–17.5%, 3.4–5.2%, and 7.5–9.5%, respectively. The agreement of Elite microplate analyzer with CODA analyzer was 99.5%.

Conclusions

The performance of Elite microplate analyzer and the related assays on analytical sensitivity, precision, early detection, diagnostic sensitivity and specificity was all adequate for a mass screening. However, further large multi-center studies should be performed to validate our results.

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Keywords: Hepatitis B virus, Hepatitis C virus, Human immunodeficiency virus, Evaluation, Sensitivity, Specificity, Elite Microplate Analyzer

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Table 1.

Low detection limit of Monolisa HBsAg ULTRA test for HBsAg WHO standard material

WHO standard (IU/mL)	1st replicate		2nd replicate		3rd replicate		4th replicate
WHO standard (IU/mL)	S/C Ratio	Result	S/C Ratio	Result	S/C Ratio	Result	S/C Ratio	Result
3.3	40.9	P	38.9	P	46.3	P	43.8	P
0.33	6.4	P	6.1	P	6.6	P	6.8	P
0.1	2.3	P	2.4	P	2.4	P	2.3	P
0.075	2.2	P	2.2	P	2.6	P	2.4	P
0.05	1.2	P	1.7	P	1.2	P	1.2	P
0.025	0.6	N	0.7	N	0.7	N	0.7	N
0.0125	0.4	N	0.5	N	0.5	N	0.5	N

Abbreviations: P, positive; N, negative; S/C, signal/cut-off ratio.

Table 2.

Precision performance of Bio-Rad ULTRA line kits on Elite microplate system

Item	HBsAg		HCV Ag-Ab			HIV Ag-Ab
level	1	2	1	2	3	1	2
Mean S/C	1.2	11.6	1.8	2.7	6.2	1.9	5.3
Within-run CV	7.2	5.0	3.8	2.5	1.5	2.0	4.5
Between-run CV	13.7	5.4	0.7	4.2	1.2	3.9	2.2
Between-day CV	8.2	11.7	3.5	2.3	2.8	6.1	8.0
Total CV (%)	17.5	13.8	5.2	5.4	3.4	7.5	9.5

Abbreviations: S/C, signal/cut-off ratio; CV, coefficient of variation.

Table 3.

Comparison of the performance of Elite microplate system, Abbott Prism and Abbott AxSYM version 2 in seroconversion panels

Panel serial No.	Days since 1st bleed	Elite microplate (S/C ratio)	Abbott Prism^* (S/C ratio)	Abbott AxSYM version 2^* (S/C ratio)
HBV BBI
PHM935A–02	2	0.1	0.5	0.4
PHM935A–03	7	0.2	0.7	0.4
PHM935A–04	9	0.2	0.3	0.4
PHM935A–05	14	0.1	0.4	0.4
PHM935A–06	16	0.2	0.6	0.4
PHM935A–07	21	1.5	1.8	0.7
HCV BBI
PHV916–02	2	0.6	0.1	0.4
PHV916–03	7	0.1	0.1	0.3
PHV916–04	9	1.0	0.2	0.4
PHV916–05	16	0.8	1.0	0.2
PHV916–06	19	2.2	1.9	4.4
PHV916–07	23	3.4	2.7	15.5
HIV BBI
PRB931–02	2	0.4	0.3	0.4
PRB931–03	7	0.4	0.4	0.3
PRB931–04	9	0.5	0.3	0.4
PRB931–05	15	0.9	0.3	0.4
PRB931–06	28	37.8	7.0	7.5
PRB931–07	33	37.8	64.9	14.2

^* Information was obtained from seroconversion panel data sheets.

Table 4.

Summary of the diagnostic sensitivity and specificity of Bio-Rad Monolisa HBsAg, HCV Ag-Ab, and Genscreen HIV Ag-Ab ULTRA kits on Elite microplate system

Performance item	Test item	Test sample number	Elite microplate		Performance (95%CI)
Performance item	Test item	Test sample number	Positive	Negative	Performance (95%CI)
Diagnostic sensitivity	HBsAg	99	99	0	100%
					(96.3–100%)
	HCV Ab	97	97	0	100%
					(96.2–100%)
	HIV Ab	112	112	0	100%
					(96.7–100%)
Diagnostic specificity	HBsAg	1,400	0	1,400	100%
					(99.7–100%)
	HCV Ab	1,670	4	1,666	99.8%
					(99.4–99.9%)
	HIV Ab	1,704	1	1,703	99.9%
					(99.7–99.9%)

Abbreviation: CI, confidence interva.

Table 5.

Comparison of anti-HCV results of LG HCV Ab 3.0 EIA test kit (LG chemicals, Seoul, Korea) performed on the different automatic analyzers, CODA system and Elite microplate system

		LG HCV Ab test on Elite system		Total	Kappa^†
		Positive	Negative	Total	Kappa^†
LG HCV Ab test on CODA system	Positive	30	6^*	36	0.88
	Negative	2^*	1,494	1,496
Total		32	1,500	1,532

^* Eight discrepant results were confirmed as all negative by RIBA test.

^† Standards for strength of agreement for the kappa coefficient: ≤0=poor, 0.01–0.20=slight, 0.21–0.40=fair, 0.41–0.60=moderate, 0.61–0.80=substantial, and 0.81–1.0=almost perfect[29].

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