1. RESEARCH ON THE PRODUCTION OF HBsAg STANDARD SERUM FOR QUALITY CONTROL
Prof. Dao Quang Minh
PD.MD. Nguyen Minh Hiên
MD. Nguyen Quang Hieu

2. PRESENT CONTENTS - Reason for study selection Objectives
Some basic issues related to Study
- Materials and methods
- Results and discussion
- Conclude

3. REASON FOR STUDY SELECTION

4. Vietnam is high HBV located
Quality Assurance
Standard Sample
Vietnam is high HBV located
HBsAg quantitative tests
RESEARCH ON THE PRODUCTION OF HBsAg STANDARD SERUM FOR QUALITY CONTROL

5. OBJECTIVES 1. Product process of HBsAg Standard Serum
2. Evaluate the quality of HBsAg Standard Serum

6. SOME BASIC ISSUES RELATED TO STUDY

7. HEPATITIS B VIRUS Antigen Structure: Dane particles HBsAg HBcAg HBeAg
HBsAg: Evidence of HBV infection

8. HBsAg DETECTION TECHNIQUES
Direct: Finding Dane particle
Indirec:
+ Elisa (Enzyme linked immunosorbent assay)
+ IEP (Immunoelectrophoresis)
+ Fluorescent Immunity (most popular method)

9. Internal Quality Control External quality control
QUALITY ASSURANCE
Internal Quality Control
External quality control
Standard Samples

10. STANDARD SAMPLES Certified Reference Materials (CRM)
Standard Samples are produced in different forms for different purposes
Requirement of Standard Samples:
Ensure homogeneity
Ensure stability over time

11. MANUFACTURING HBsAg STANDARD SERUM (HSS)
CDC (USA): Producing and supplying the dry plasma for many countries.
Standard laboratory (Australia, England, France, Korea, Thailand) produced HBsAg Standard Serum from serum or plasma.
According to ISO/IEC 17043:2011 and TCVN 7366, TCVN 8245

12. MANUFACTURING HBsAg STANDARD
In Vietnam:
Mainly importing standard samples from Roche, Beckmancounter, Randox ...
High cost
Not be active because of dependence on imports (including the licences)

13. MATERIAL AND METHOD

14. MATERIAL OF STUDY
* Plasma samples that HBsAg were positive at National Institute hematology and blood transfusion, Viet Duc Hospital The Selections: HBsAg is positive, The samples are not hemolytic, Precipitation. * The other
name
Manufacturer
Made in
HBsAg II
Roche
USA
HBsAg QN
Proclin
Supelco
Formalin
Qingdao
China
CaCl2 anhydrous
(NH4)2SO4 anhydrous

15. Time: From 3/2016 to 11/2016
Study Location:
Quality control center for medical laboratory in HMU
Biochemistry of Department in Thanh Nhan Hospital
Method
Production testing

16. BUILDING HBsAg STANDARD SAMPLES
Samples have two levels:
Level 1: [HBsAg]: During 7,500 – 11,500 IU/mL
Level 2: [HBsAg] During 35 – 60 IU/mL
Sample has been preserved two ways
at-300C
at 00C
Time for preservation: 150 days

18. RESULT AND DISCUSSION

19. RESULTS OF COLLECTION Collections Volume [ HBsAg ] COI Sample 1 100 ml
3857
Sample 2
1631
Sample 3
2525
Sample 4
1143
Sample 5
1674
Total sample
500ml
4850

20. RESULT OF CHANGED FROM PLASMA TO SERUM
* H.T.T.Hà (2014): 0,01M
* Jin-Sook Wang (2012): 0,01M
* WHO/UNAIDS (1996)
Number
Calcium concentration
(M)
Volume
Before
(ml)
[HBsAG]
(COI)
after 1
0,1 10
4850
2,5
5072 2
0,05
5,0
5019 3
0,01
7,5
5025
Remain
470
352
5021

21. [ HBsAg]: CONCENTRATE AND DILUTION
Original
Concentrate
Dilution
200 times
Result
(IU/mL)
2101
11588
56,92

22. EVALUATION OF HOMOGENEITY HSS
Samples
Result (IU/mL)
Level 1
Level 2
time 1th
time 2th
Sample 1
11337
11134
56,58
55,17
Sample 2
11521
11057
56,99
55,40
Sample 3
11255
11726
56,29
55,37
Sample 4
11163
11359
56,06
56,66
Sample 5
11227
11686
55,62
57,22
Sample 6
11652
11570
56,27
55,77
Sample 7
11498
11603
55,88
55,95
Sample 8
11320
11606
56,54
55,81
Sample 9
11117
11111
55,90
Sample 10
11187
11157
56,28
56,64
Mean
11327,7
11400,9
56,31
55,99 SE
55,6
83,8
0,124
0,207 SD
175,9
265,3
0,39
0,66
CV(%)
1,55
2,32
0,69
1,17
P (Ttest)
0,45
0,32
Roche Diagnotics: 5,6%
Abbot Diagnotics: 6,7%

23. EVALUATION OF STABILITY OF LEVEL 1 HSS
Time
date
Result (IU/mL)
0oC
-30oC
time 1th
time 2th 1
29/6/2016
11078
11107
11274
11250 2
11/7/2016
11100
11190
11390
11500 3
20/7/2016
11140
11194
11494
11411 4
29/7/2016
11184
11128
11088 5
9/8/2016
11685
11439
11334 6
18/8/2016
11697
11653
11553
11519 7
28/8/2016
11299
11214
11114
11414 8
7/9/2016
11075
11023 9
17/9/2016
11186
11109
11409
11352 10
27/9/2016
11156
11197
11154 11
7/10/2016
11044
11138 12
17/10/2016
11592
11410
11289
11248 13
27/10/2016
11499
11562
11262
11322 14
6/11/2016
11364
11393
11493
11613 15
16/11/2016
11498
11464
11423
mean
11306,5
11281,4
11308,5
11329,1 SD
231,7
189,4
168,9
154,4 SE
59,8
48,9
43,6
39,9
CV(%)
2,05%
1,68%
1,49
1,36
P (Ttest)
0,23
0,37
Roche Diagnotics: 5,6%
Abbot Diagnotics: 6,7%

24. EVALUATION OF STABILITY OF LEVEL 1 HSS
AT -300 C VÀ 00C

25. EVALUATION OF STABILITY OF LEVEL 2 HSS
Time
date
Result (IU/mL)
0oC
-30oC
time 1th
time 2th 1
29/6/2016
55,08
55,63
53,51 54 2
11/7/2016
55,78
56,01
56,15
55,76 3
20/7/2016
55,31
54,93 4
29/7/2016
55,82
55,90
55,37
55,10 5
9/8/2016
54,99
55,29
55,49
55,07 6
18/8/2016
55,91
56,12
55,62
54,97 7
28/8/2016
53,96
54,78
55,71 8
7/9/2016
54,11
54,66
53,92
54,28 9
17/9/2016
55,67
55,73
56,63
55,99 10
27/9/2016
55,40
56,00
54,47
55,23 11
7/10/2016
55,09
54,95
56,5
56,19 12
17/10/2016
55,14
55,26 55 13
27/10/2016
55,83
55,11 14
6/11/2016
55,22
54,89
55,35
55,92 15
16/11/2016
54,61
55,00
55,96
55,13
Mean
55,44
55,32 SD
0,57
0,51
0,52
0,86 SE
0,148
0,132
0,133
0,22
CV(%)
1,03
1,04
0,93
1,56
p (T test)
0,34
0,11
BeckmanCounter: 8,3%
Abbot Diagnotics: 7,8%

26. EVALUATION OF STABILITY OF LEVEL 2 HSS
AT -300 C VÀ 00C

27. RESULT OF THE CULTURE Date At 370C At 220C tube 1 tube 2 tube 3 tube 4
(-)
7th
14th

28. CONCLUSION - HBsAg standard serum test has been developed .
- Homogeneity and stability in 5 months at 0oC and -30oC According to the TCVN 8245, ISO Standard.