Q PROCEDURE FORMAT - Pusat Pengajian Sains … · PROCEDURE NO. HUSM/ENDO/STM 4 ... Automated Cobas...

ENDOCRINE LABORATORY, HOSPITAL UNIVERSITI SAINS MALAYSIA

TITLE : HCG + β

(Intact human chorionic gonadotropin + the β-subunit)

VERSION NO. 1

PROCEDURE NO. HUSM/ENDO/STM 4 VERSION DATE. 17.09.2014

APPROVED BY:

……….…………………………………..

DR WAN MOHD IZANI WAN MOHAMED ENDOCRINE LABORATORY DIRECTOR

CONTROLLED COPY NO: 0

REGISTERED HOLDER

ENDOCRINOLOGY LABORATORY

PREPARED BY : MUHAMMAD AMALI BIN KAMARUDIN DESIGNATION : QUALITY MANAGER / SCIENCE OFFICER : AINUN IZZATI RAZALI :DOCUMENT CONTROLLER :AFIQAH IYLIA AB HALIM :DEPUTY DOCUMENT CONTROLLER CHECKED BY : MUHAMMAD AMALI BIN KAMARUDIN DESIGNATION : ENDOCRINE LABORATORY DEPUTY DIRECTOR/QUALITY MANAGER AUTHORISED BY : DR WAN MOHD IZANI BIN WAN MOHAMED DESIGNATION : ENDOCRINE LABORATORY DIRECTOR

BHCG (Intact human chorionic gonadotropin + the β-subunit)

of 14 STANDARD TECHNICAL MANUAL


TITLE : HCG + β


VERSION NO. 1


1. CONTENT

NO. CONTENT PAGE

1. Content 2

2. Record of Review 3

3. Record of Amendments 4

4. Objective, method, test principle 5

5. Clinical significance 6

6. Requirements 7-8

7. Procedure 9-13

8. Limitation / source of error 14

9. Limits and Ranges 14

10. References 14



TITLE : HCG + β


VERSION NO. 1


2. RECORD OF REVIEW

No

semakan

Tarikh

semakan

Nama Penyemak Tandatangan

pengesahan

1 22/01/2014 Muhammad Amali b Kamarudin

2 17/09/2014 Ainun Izzati Razali



TITLE : HCG + β


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3. RECORD OF AMMENDMENT DATE VERSION

NO DETAIL OF AMMENDMENT BY

17/09/2014 1.0 9.4.3 Procedure dispatch of results

Remove:

a.original request forms and printed will be not

dispatched accept for STAT sample will be

manually dispatched via Endocrine laboratory

pigeonhole or direct to wad/ clinic.

b.Carbonized copy of request form and its printed

results for the day is compiles the next day and

kept for 3 years.

c.Results are also kept in LIS database for

traceability.

Izzati



TITLE : HCG + β


VERSION NO. 1

PROCEDURE NO. HUSM/ENDO/STM 4 VERSION DATE. 17.09.2014 4. OBJECTIVE

Standard technical manual to determine in vitro quatitative of the sum of human chorionic

gonadotropin (hCG) plus the hCG β-subunit in human serum and plasma on an

Automated Cobas e Immunoassay Analyzers.

• Early detection and monitoring of pregnancy

• To evaluate the risk of trisomy 21 (Down Syndrome)

• To serve the management of patients with trophoblastic disease.

5. METHOD

In vitro quantitative determination of the sum of human chorionic gonadotropin (hCG) plus

the hCG β-subunit in human serum and plasma are performed using Automated Cobas e

Immunoassay Analyzers based on electrochemiluminescent technology that is the using

of ruthenium complex and the measuring cell.

6. TEST PRINCIPLE

Sandwich principle. Total duration of assay: 18 minutes.

• 1st incubation: 10 µL of sample, biotinylated monoclonal hCG-specific antibodies,

and a monoclomal hCG-specific antibody labeled with a ruthenium complex

[Tris(2,2’-bipyridyl)ruthenium(II)-complex (Ru(bpy)3 2+) react to form a sandwich

complex.

• 2nd incubation: After addition of streptavidin-coated microparticles, the complex

becomes bound to the solid phase via interaction of biotin and streptavidin.

• The reaction mixture is aspirated into the measuring cell where the microparticles

are magnetically captures onto the surface of electrode. Unbound substances are

then removed with ProCell. Application of a voltage to the electrode then induces

chemiluminescent emission which is measured by a photomultiplier.

• Results are determined via a calibaration curve which is instrument-specifically

generated by 2-point calibration and a master curve provided via the reagent

barcode.



TITLE : HCG + β


VERSION NO. 1

PROCEDURE NO. HUSM/ENDO/STM 4 VERSION DATE. 17.09.2014 7. CLINICAL SIGNIFICANCE

HCG is produced in the placenta during pregnancy. In non pregnant women, it can also

be produced by tumors of the trophoblast, germ cell tumors with trophoblastic components

and some non-trophoblastic tumors. Human chorionic gonadotropin consists of a number

of isohormones with differing molecular size. The biological action of hCG serves to

maintain the corpus luteum during pregnancy. It also influences steroid production. The

serum of pregnant women contains mainly intact hCG. Elevated values here serves asa

an indication of chorionic carcinoma, hydatiform mole or multiple pregnancy and

depressed values indicate threatening or missed abortion, ectopic pregnancy, gestosis or

intra-uterine death.

Measurement of Hcg+β makes also a contribution to risk assessment for trisomy 21

(Down Syndrome) in the second trimester of pregnancy together with AFP and others

paramenters, such as exact gestational age and maternal weight.



TITLE : HCG + β


VERSION NO. 1

PROCEDURE NO. HUSM/ENDO/STM 4 VERSION DATE. 17.09.2014 8. REQUIREMENTS

8.1 EQUIPMENT Cobas e 411/601 Analyzer, Centrifuge, vortex mixer.

8.2 REAGENT

REAGENT CAPACITY

PER CONTAINER

CODE NUMBER

STORAGE (ºC)

M Streptavidin-coated microparticles (transparent cap) 1 bottle, 6.5 mL M 2-8 R1 Anti-hCG-Ab~ biotin (gray cap)

1 bottle, 9 mL R1 2-8

R2 Anti-hCG-Ab~Ru(bpy)32+ (black cap) R2 2-8

ProCell M system buffer 2 X 2 L

04880340190 2-8 CleanCell M measuring cell cleaning solution 04880293190 2-8 ProbeWash M cleaning solution for run finalization and rinsing during reagent change 12 X 70mL 03005712190 2-8

Elecsys SysClean system cleaning solution 5 X 100mL 11298500316 2-8 PreClean M cleaning solution for run finalization and rinsing during reagent change 5 X 600mL 03004899190 2-8

REAGENT CAPACITY

PER CONTAINER

CODE NUMBER

STORAGE (ºC)

SysClean Adapter M - 03027651001 2-8

8.1.2 SHELF LIFE (days)

Unopened at 2-8 ºC Up to the stated expiration date

After opening at 2-8 ºC 84 days

On the analyzer 56 days

8.3 QUALITY CONTROL MATERIAL

QC NAME CAPACITY PER VIAL

Lyphochek Immunoassay Plus Control

Level 1 Level 2 12 X 5 mL Level 3

8.4 SPECIMEN 8.4.1 Serum collected - using standard sampling tubes or tubes containing

separating gel. Li-, Na-, NH+4-HEPARIN, K3-, Na2-EDTA, sodium citrate plasma,

Plain Tube, sodium fluoride/potassium oxalate plasma.

8.4.2 Sample preparation - a. Draw 3 ml blood from the patient and transfer it into gel tube

b. Leave it at room temperature not less than half an hour.

c. Centrifuge at 3000 – 3500 rpm for 5 minutes.



TITLE : HCG + β


VERSION NO. 1

PROCEDURE NO. HUSM/ENDO/STM 4 VERSION DATE. 17.09.2014 d. Transfer the supernatant (serum) as much as possible into 5 ml polypropylene

tube.

e. Run the test or keep it in the fridge or freezer. (Stability: 3 days at 2 – 8 C, 12

months at -20 C)

Ensure the patients’ samples, calibrators and controls are at ambient temperature (20 – 25 C) before

measurement because of possible evaporation effects, samples, calibrators and controls on the analyzers

should be measured within 2 hours.



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9 PROCEDURE

NO. ACTIVITY RESPONSIBILITY 9.1 START UP

9.1.1 Pre-Operational Checks

a. Refill printer with printing paper. b. Check solid waste and liquid waste (if required). c. Check reagent supply and expiry dates (replace if required).

9.1.2 Start Up System When the system is in sleep mode, it starts up automatically by using sleep

pipe function (power up pipe with starting time)

a. Turn ON instrument. When the automatic power up is specified in the system, the pipe function will be executed automatically.

b. Switch on water supply. c. Swith on control unit computer. d. Logon to the system using your operator ID and password.

[Username: user, password : ENDO ]

MLT/SO

9.1.3 Perform Daily Check up

a. Check system alarm b. Execute maintenance action or maintenance pipe function.

9.1.4 Perform Sample Data Clear All measurement results of routine and STAT samples will be deleted. The

measurement results of quality control samples will be moved to the QC view area of data storage control section.

a. Go to sample data clear. b. Choose clear or backup and clear. c. Click ok.

9.1.5 Perform Reagent Preparing

a. Active the Preventive Action check box on the System Overview screen.

b. Reagent Load List will be printed for reagent with yellow, purple or red

alarm. c. Load reagents according to reagent load list onto the instrument. d. Remove all empty packs from the system.

e. Perform Prime for ISE. Select all. f. Execute after the Reagent Prime for ISE, recalibrate the ISE unit

before you resume routine operation.

Daily Maintenance Select Execute

Sample Data Clear Clear or Backup and Clear OK

Reagent Preparing Reagent Load List YES

Reagent Preparing Reagent Prime



TITLE : HCG + β


VERSION NO. 1


NO. ACTIVITY RESPONSIBILITY 9.2 CALIBRATION AND QUALITY CONTROL

9.2.1 Setup New Internal Quality Control Lot The system automatically recommends calibrations and controls to be

performed. But the recommended calibrations and QCs must be confirmed by the operator before they can be measured.

a. Print the Calibrator and QC Load List

b. Load calibrator and QC racks on the analyzer according to calibrator

and QC load list.

9.2.2 Internal Quality Control Analysis CONTROLS (Lyphochek Immunoassay Plus Control Levels 1, 2 and 3)

a. Reconstitute each vial with 5.0 mL of distilled or deionized water by using volumetric pipet.

b. Replace the stopper and allow the control to stand for 15 minutes. c. Before sampling, allow the control to room temperature and swirl gently

to ensure homogeneity. d. Aliquot controls (250 uL aliquot volume) separately into hitachi cup for

use. Note for e 601: e pack should be placed on board before, because lot specific target values can be coded within the reagent barcode.

e. Select one of the available search conditions.

f. Activate the check box of the parameter to be download

g. Check, if control information is downloaded.

MLT/SO 9.2.3 SET UP CONTROLS MANUALLY

a. Select one of the available search conditions. b. Select a blank line

c. Type all necessary data in the Control Window.

The new control appears on the control list.

d. Enter target mean, target SD, and confidence limits High and Low, if

necessary. 9.2.4 ACTIVATING CONTROL TESTS

a. Select control name and test to be activated.

b. Activated test highlights green.

Calibration and QC select Recommended

Print

Start

QC Install Download

Search

Download OK OK Close

QC Install

Add

OK

Edit

QC Install

Active Test



TITLE : HCG + β


VERSION NO. 1


NO. ACTIVITY RESPONSIBILITY 9.2.5 ASSIGN RACK FOR NON BARCODED CALIBRATORS

MLT/SO

c. Select the calibrator to be assigned to a rack ID and position from the

list on the left to the list on the right.

d. To assign the selected calibrator to the selected position.

e. To undo the assignment. f. After all assignment are completed

Note : Do not mix barcode and non-barcoded calibrator/control on the same rack. 9.2.5 ASSIGN RACK FOR NON BARCODED CONTROL

a. Select the control from the list on the left and an unsigned rack number

and position from the list on the right.

b. To assign the selected control to the selected position. c. To undo the assignment.

d. After all assignment are completed

Note : Do not mix barcode and non-barcoded calibrator/control on the same rack.

9.2.6 CHECK THE UNIT OF ANALYTE IN THE ANALYZER

a. To check the Unit of the analyte used on the analyzer,click on:

MLT/SO

9.3 SAMPLE ANALYSIS 9.3.1 Sampler Cobas e411/601

a) Make sure that all instrument in STANBY mode, QC PASSED b) Make sure LIS connection is ONLINE and LIVE.

MLT/SO

Calibration Calibrator Rack Assign

Assign ------>

<------- Remove

OK

QC Control Rack Assignment

Add ------>

<------- Remove

OK

Utility Application Range



TITLE : HCG + β


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NO. ACTIVITY RESPONSIBILITY

c) Place the samples in the cassette properly.

d) Load the cassette on the sample loader and press START.

e) For STAT sample tube, load the sample at the STAT station

f) Discard and keep the sample from the cassette or the station once the

analysis completed at a sample rack.



TITLE : HCG + β


VERSION NO. 1


NO. ACTIVITY RESPONSIBILITY

For manually: a) Go to

b) Enter sample ID (Refer to registration barcode sample)

c) Enter rack number and position. d) Double check sample ID.

e) Put sample at the single tube presentation station. f) Then finally push.

9.4 RESULT REPORTING 9.4.1 Printing of results

a. Test results will be automatically printed. b. Staple printed results accordingly to request forms.

9.4.2 Validating & Verification

a. Validate the results. b. Verify the transcription of results in the LIS.

MLT/SO

9.4.3 Dispatching of Results

a. Release the results in LIS, put comment if required. b. Results are also kept in LIS database for traceability.

MLT/SO

Workplay Test Selection

Enter Select Test Barcode Read Error

Add OK Save

Start



TITLE : HCG + β


VERSION NO. 1


10 LIMITATION/SOURCES OF ERROR 10.1 In patients receiving therapy with high biotin doses (>5 mg/day), no sample should

be taken until at least 8 hours after the last biotin administration. 10.2 Extremely high titers of antibodies to analyte-specific antibodies, streptavidin or ruthenium can occur and will affect hCG result. 10.3 Lysed blood sample.

11 LIMITS AND RANGES

11.1 Measuring Range

0.100 mIU/mL – 10000 mIU/mL

11.2 Reference Range

Non pregnant (pre menopausal female) : < 1 IU/L Post menopausal: Female < 7 : Male < 2

12 REFERENCES 9.1 Cobas 6000 analyzer series Operator’s Manual. 9.2 Cobas 4000 analyzer series Operator’s Manual. 9.3 Elecsys and cobas e analyzers reagents for HCG+β (Package Insert) 9.4 Bio-Rad Lyphochek Immunoassay Plus Control Level 1, 2, and 3 (Package Insert)

End of Document


Q PROCEDURE FORMAT - Pusat Pengajian Sains … · PROCEDURE NO. HUSM/ENDO/STM 4 ... Automated Cobas...

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