Prof. Musabaev E.I.

Quality Control Systems Quality Control Systems for Hepatitis B and C for Hepatitis B and C

diagnostics in diagnostics in Uzbekistan. Uzbekistan.

Prevalence of HBV and HCV infections in UzbekistanPrevalence of HBV and HCV infections in Uzbekistan

Examined group Quantity

of examined peopleHBsAg

%anti-HCV

%

Blood-donors 2030 4,8 9,0

Contacted to viral hepatitis

556 15,1 0,9

Pregnant 731 16,4 1,1

Drug addict 115 6,0 11,3

Viral hepatitis - patients

1212 20,5 3,7

Others 62 17,1 4,8Total 4706 12,0 5,6

Background

Background

February, 1998. The agreement amongst Government of Swiss Confederation, Division of Humanitarian Aid and Swiss Disaster Relief (SDR) and Republic of Uzbekistan "Aid in the fight against viral hepatitis in Uzbekistan"

March, 1999. Establishing of the Reference Laboratory Autumn, 1999. Educational program for laboratory specialists of

Uzbekistan was developed in common with Centers for Disease Control and Prevention (CDC, USA).

Autumn, 1999. First reference HBsAg panel was created with specialists of CDC and Swiss Quality Control center.

November, 1999. First Qulity Control Seminar for laboratory specialists of Uzbekistan organised by CDC.

October, 2000. Mandatory of participation in EQA

DatesDates

Creation of panels for EQA for the following markers (concerning the priorities):

• HBsAg

• anti-HCV

• AIDS/anti-HIV-1 and anti-HIV-2 Providing of Diagnostic laboratories of the Republic by the reference panels

created in Reference laboratory.

External Quality Assessment of the Diagnostic laboratories of:• blood banks;• sanitary-epidemiological stations;• infectious diseases hospitals;• private diagnostic clinics;

Aims & Tasks

Improvement of Efficiency and Effectiveness of Improvement of Efficiency and Effectiveness of Diagnostic LaboratoriesDiagnostic Laboratories

Quality Control of imported and home-produced ELISA test-systems with a purpose of preparation/taking decision about possibility of their licensing and further using.

• Conducting the trials of diagnostic kits with use of Reference panels and/or and 200-250 serum samples received from different regions of the Republic attested with Referent ELISA test-system (Ortho Diagnostic Systems);

Rendering methodical and technical aid to the home-producers of ELISA.

• Trials of kits during its construction;

• Interpretation of results;

Improvement of Diagnostic Kits Improvement of Diagnostic Kits Quality Quality Used in Used in UzbekistanUzbekistan

Aims & Tasks

Training specialists of diagnostic laboratories of Uzbekistan in essential theoretical and practical skills:

• Development of the educational programs for the low- and middle-level specialists of diagnostic laboratories;

• Organisation and conducting practical symposia for epidemiologists in the different regions of Uzbekistan

• Organisation and conducting practical symposia for diagnostic laboratory specialists

Raising the Education Level of Medical SpecialistsRaising the Education Level of Medical Specialists

Aims & Tasks

№1 - 4000 ng - native highly reactive serum

№2 - 4 ng - 1000-fold dilution of serum №1

№3 - 1 ng - 4000-fold dilution of serum №1

№4 - 0,5 ng - 8000-fold dilution of serum №1

№5 - 0,25 ng - 16000-fold dilution of serum №1

№6 - 0,125 ng - 32000-fold dilution of serum №1

№7 - Diluent pool

№8 - Serum negative for HBsAg HBsAg concentration is determined by calibration against ay-subtype of HBsAg standard

presented by CDC (USA).

Calibration was carried out only for the samples №2-6 whose reactivity was directly proportional to the dilution degree (Lambert-Ber low).

Calibration was performed with “Ortho” (USA) and “Dia-Sorin” (Italy) ELISA

HBsAg-panel for EQCHBsAg-panel for EQC

Results

Anti-HCV panel for EQCAnti-HCV panel for EQC

Results

Table 5. Reactivity and antibody profiles of anti-HCV panel’s specimens

RIBA HCV 3.0 SIAOrtho 3.0 HCVantigen band patternsSample

№ S/CO c100 c33c c22 NS51 pos 9,77 2+ 3+ 4+ 4+2 pos 7,90 +/- 1+ 4+ 2+3 pos 3,82 - +/- 4+ +/-4 pos 1,44 - +/- 3+ +/-5 neg 0,406 pos 2,17 +/- +/- 2+ -7 pos 1,99 - 3+ 3+ -8 neg 0,12

Questionnaire for External Quality Questionnaire for External Quality AssessmentAssessmentQuestions for External Quality Control of Diagnostic Labs in Uzbekistan

Name of laboratory––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––

Date –––––––––––––––

StuffDoctorsTechniciansLaboratory assistantsLaboratory premises

Boxes Preparatory rooms Rest rooms Other premises

Laboratory equipment Laboratory furniture (tables, with treatable surface) Reader for estimation of ELISA Refrigerator (4 - 6 С ), presence of thermometer, regularity of

temperature recordings Refrigerator (-20 С) Distillator Thermostat, presence of thermometer, regularity of temperature

recordings Calculator Timer Dry-heating safe

Automatic pipettes 10 l 20 l 50 l 100 l 500 l 1000 l Multi-channel pipettes Filter paper

Laboratory glassware kits Petry plates Flasks for dilution Graduated cylinders and glasses Pipette tips

Laboratory safety Gloves Presence of disinfectant Presence of 6% hydrogen peroxide (for treatment of utilised pipette tips) Presence of auto-clave

Sort and volume of analyses per month Hepatitis А markers Hepatitis В markers Hepatitis С markers HIV markers Others

Documentation of receiving, registration and storing of specimens

Unified form of assignment of specimens to a laboratory Unified form of registration of specimens Conforming to the rules of storage of blood samples Conforming to the rules of serum isolation Conforming to the rules of marking and registration of specimens Conforming to the rules of storage of serum samples

Соблюдение условий постановки ИФА Presence of instruction for ELISA Presence of experiment protocols Presence of reference standards Correctness of registration of results Correctness of protocol composing Presence of unified form for registration of results Presence of unified form of answer

Training Improvement of professional skill of stuff Teaching people from other laboratories

Results

Dilutions of positive (12/20) Negative (6/6)

10 18 35 119 162

125

128

133

144

120

Diagnostic Lab

U 16 64 128

512

U 16

64

128

256

U 16

64

128

256

U 16

64

128

256

Number ofsample

Effectivenessof laboratory

tests

№

City Organization

Reagents

1 2 3 4 5 6 7 8 9 10

11

12

13

14

15

16

17

18

19

20

21

22

23

24

25

26

Sensibility Specificity

ReferenceLab

Ortho + + + + - + + - - - + + - - - + + + + - - - - - - - 100% 100%

Pasteur Inst. + + + - - + + - - - + + - - - + + - - - - - - - - - 75% 100%1 Obl. SES Posbon + + - - - + - - - - + + - - - + + - - - - - - - - - 58% 100%2 City SES Posbon + + + + - + - - - - + - - - - + + - - - - - - - - - 67% 100%

Posbon + + - - - + - - - - + - - - - + + - - - - - - - - - 50% 100%3 TashkentCity BTC Vector Best + + + + - + - - - - + + - - - + + + - - - - - - - - 83% 100%

4 SES Posbon + + + - - + - - - - + - - - - + + - - - - - - - - - 58% 100%5

Navoi

Obl.BTCi - -6 Obl. SES Posbon + + + - - + - - - - + - - - - + + - - - - - - - - - 58% 100%7

Bukhara

Obl.BTC Posbon + + + - - + - - - - + + - - - + + - - - - - - - - - 67% 100%8 Obl. SES Vector Best + + + + + + + - - - + + + + - + + + + - - - - - - - 100% 79%9

Samarkand Obl. BTCii Posbon + + - - - + - - - - + - - - - + - - - - - - - - - - 42% 100%

i No reagent availableii No instrument working, only visual measurement

Results of External Quality Control of HCV diagnostic in some provinces of Uzbekistan

Results

National Reference HBsAg panel for quality control of diagnostic kits

HBsAg positive samples (N=9) are diluted to the end point in the negative for all viral hepatitis markers human serum pool (N=2). HBsAg negative samples (N=36) are undiluted.

Titration: 1:1; 1:10 000; 1:100 000; 1:200 000; 1:500 000;

0.052Blanc 0.172 3.509 0.176 1.278 0.070 0.133 0.061 0.081 0.071 0.069

0.064Nc 0.124 1.354 0.063 0.178 3.113 0.072 0.069 0.059 0.062 0.071

0.065Nc 0.077 0.354 3.073 0.078 0.706 0.070 0.065 0.068 0.076 0.083

0.060Nc 3.477 0.103 1.186 0.069 0.206 0.064 0.068 0.081 0.066 0.065

1.586Pc 1.180 0.079 0.186 3.243 0.102 0.067 0.065 0.068 0.067 0.087

1.610Pc 0.208 3.384 0.080 0.851 0.070 0.076 0.085 0.066 0.065 0.069

3.482 0.112 1.940 0.079 0.183 3.306 0.060 0.068 0.065 0.058 0.069

1.248 0.077 0.194 3.345 0.090 0.769 0.065 0.072 0.062 0.059

Cutoff=0.092Results of attestation of control panel performed with ELISA "ORTHO DIAGNOSTIC SYSTEMS"

Results

Field of Our ActivityField of Our Activity

Sanitary Epidemiological Control

Services14 labs

Manufactures of Diagnostic kits:

Nihol, Uzbekistan;Posbon, Uzbekistan;

Radiopreparat, Uzbekistan;Institute of Genetics, Uzbekistan;

Vector Best, Russia

Private Diagnostic Laboratories

Regional Blood Transfusion Services

15 labs

Framework

Regional AIDS Centres

15 labs

The founders of the Reference laboratoryThe founders of the Reference laboratory

SWISS AGENCY FOR DEVELOPMENT COOPERATIONFederal Office for Foreign Economic Affairs

MINISTRY OF PUBLIC HEALTH

SERVICES OF UZBEKISTAN

SWISS QUALITY CONTROL CENTERGeneva, Switzerland

Framework

CDC, International and viral hepatitis Department, Atlanta, USA

Department of Virology, University of Helsinki, Finland Tarasevich’s Institute of Standardisation and Quality Control,

Moscow, Russia Departament of blood tranfusion,University of Nogoi,Japan National Reference Laboratory of Kirgiztan National Reference Laboratory of Turkmenistan National Reference Laboratory of Kazahstan

CollaborationCollaboration

Framework

Acknowledgements

This work was supported by Swiss Disaster Relief (SDR), Swiss Agency for Development and Cooperation and Centers for

Disease Control and Prevention (CDC, USA) Personal thanks to: Dr. Favorov M.O. Regional Director of CDC’s

programms in Central Asia Dr. Deom A. Director of Swiss Quality Control

Center Dr. Kalashnikova T.V. CDC, consultant

Reference laboratory staff:Ongarbaev A.Mustafaeva E.Albetkova A.

Acknowledgements