1. طرق التحليل المناعي للكشف عن السموم والمؤثرات العقلية
مركز مراقبة السموم والكيمياء الطبية الشرعية بالرياض
Poison Control & Medical forensic Chemistry Center, Riyadh
طرق التحليل المناعي للكشف
عن السموم والمؤثرات العقلية
Bader A. Alnafisah
Toxicologist
Director of Poison Control
& Medical Forensic Chemistry Center, 
Riyadh KSA
Al-nafisah, B. PCC, Riyadh(2015)

2. Techniques used in Toxicology
مركز مراقبة السموم والكيمياء الطبية الشرعية بالرياض
Poison Control & Medical forensic Chemistry Center, Riyadh
Techniques used
in Toxicology
Immunoassays
Chromatographic (TLC-LC–GC-IC )
Spectroscopic ( MS-A.A-FTIR-UV)
Al-nafisah, B. PCC, Riyadh(2015)

3. مركز مراقبة السموم والكيمياء الطبية الشرعية بالرياض
Poison Control & Medical forensic Chemistry Center, Riyadh
Al-nafisah, B. PCC, Riyadh(2015)

4. مركز مراقبة السموم والكيمياء الطبية الشرعية بالرياض
Poison Control & Medical forensic Chemistry Center, Riyadh
Immunoassays
chromatographic methods were employed, which had good specificity and sensitivity but were too labor intensive.
The advent of immunochemical assays, along with automation, meant faster, less labor intensive and more reliable for TDM and screening tests (not confirmatory).
Al-nafisah, B. PCC, Riyadh(2015)

5. مركز مراقبة السموم والكيمياء الطبية الشرعية بالرياض
Poison Control & Medical forensic Chemistry Center, Riyadh
Immunoassays principle are depends on the reaction of an antigen (analyte) and an antibody
All immunoassays require the use of labeled material in order to measure the amount of antigen or antibody.
Al-nafisah, B. PCC, Riyadh(2015)

6. مركز مراقبة السموم والكيمياء الطبية الشرعية بالرياض
Poison Control & Medical forensic Chemistry Center, Riyadh
Labeled Immunoassays
Antigen or antibody is labeled ( tagged ) with a substance that can be detected later on and allows for the detection of an antibody – antigen reaction
Types of tags
Radioactive isotopes
Enzymes
Fluorescent molecules
Luminescent labels
Types of reactions
Competitive binding
Noncompetitive binding
Al-nafisah, B. PCC, Riyadh(2015)

7. Separation Techniques
مركز مراقبة السموم والكيمياء الطبية الشرعية بالرياض
Poison Control & Medical forensic Chemistry Center, Riyadh
Separation Techniques
Heterogeneous assay
Bound and free antibody must be separated before label is measured
Example: ELISA (Enzyme Linked Immunosorbant Assay)
Homogeneous assay
Bound and free antibody do not need to be separated prior to measurement phase.
Example: EMIT (Enzyme Multiplied Immunoassay Technique )
Al-nafisah, B. PCC, Riyadh(2015)

8. Competitive Labeled Immunoassays (RIA, FIA, EIA)
مركز مراقبة السموم والكيمياء الطبية الشرعية بالرياض
Poison Control & Medical forensic Chemistry Center, Riyadh
Competitive Labeled Immunoassays (RIA, FIA, EIA)
A competition between tagged antigens ( reagent ) and
untagged antigens ( patient )for a limited amount of antibody ( reagent )
Al-nafisah, B. PCC, Riyadh(2015)

9. مركز مراقبة السموم والكيمياء الطبية الشرعية بالرياض
Poison Control & Medical forensic Chemistry Center, Riyadh
Immunoassays
EMIT: The EMIT (Enzyme Multiplied Immunoassay Technique)
Homogeneous (Bound and free antibody do not need to be
separated prior to measurement phase)
The EMIT (is qualitative and semi-quantitative technology) is based on competition for the target analyte antibody binding sites. Analyte in the sample competes with the drug in the enzyme reagent that is labeled with G6PDH. Active enzyme G6PDH converts the coenzyme (NAD) in the antibody reagent to NADH, resulting in a kinetic absorbance change that is measured photometrically.
Al-nafisah, B. PCC, Riyadh(2015)

10. Viva & Viva-E & Viva Twin
مركز مراقبة السموم والكيمياء الطبية الشرعية بالرياض
Poison Control & Medical forensic Chemistry Center, Riyadh
Viva & Viva-E & Viva Twin
Made by Siemens
Bench top
Relatively small
Measures multiple therapeutic drugs quantitatively
Also measures major drugs of abuse DOA)
Requires little maintenance
Flexible (Easy to use)
Applies EMIT method for all measurements
Sensitivity is not sufficient; many interference and falls positive (Disadvantages !!!)
VIVA E
Al-nafisah, B. PCC, Riyadh(2015)

11. FPIA: Fluorescence Polarization Immunoassay
مركز مراقبة السموم والكيمياء الطبية الشرعية بالرياض
Poison Control & Medical forensic Chemistry Center, Riyadh
FPIA: Fluorescence Polarization Immunoassay
This method uses a fluorescent molecule as the label instead of an enzyme, making it more sensitive. In FPIA, the patient sample is incubated with a known quantity of the fluorescent-labeled drug and an antibody specific for the drug. As in EMIT, the labeled and unlabeled drugs compete for the binding sites of the antibody. Polarized light is emitted in certain angles depending on whether the fluorescent-labeled drug is bound to antibodies or not. Since this is a competitive assay, the greater the amount of drug in the sample, the lower the amount of fluorescence.
Al-nafisah, B. PCC, Riyadh(2015)

12. AxSYM Made by Abbott Big instrument
مركز مراقبة السموم والكيمياء الطبية الشرعية بالرياض
Poison Control & Medical forensic Chemistry Center, Riyadh
AxSYM
Made by Abbott
Big instrument
Measures multiple therapeutic drugs quantitatively
Also measures major drugs of abuse DOA)
Easy to use
Requires daily maintenance
Great sensitivity
FPIA is the main technique for most TDM measurements
Relatively slow
Requires continuous manual loading of RVs when running many samples. High cost of Kit (Disadvantages !!!)
Al-nafisah, B. PCC, Riyadh(2015)

13. Chemiluminescent microparticle immunoassay (CMIA)
مركز مراقبة السموم والكيمياء الطبية الشرعية بالرياض
Poison Control & Medical forensic Chemistry Center, Riyadh
Chemiluminescent microparticle immunoassay (CMIA)
Chemiluminescence: This is a chemical reaction that emits energy in the form of light. When used in combination with immunoassay technology, the light produced by the reaction indicates the amount of analyte in a sample
label is a molecule that will react as a part of the assay, so a change in signal can be measured in the urine after added reagent solution. CMIA is non-competitive sandwich assay technology to measure analytes. The amount of signal is directly proportional to the amount of analyte present in the sample
Al-nafisah, B. PCC, Riyadh(2015)

14. CMIA is non-competitive sandwich assay technology
مركز مراقبة السموم والكيمياء الطبية الشرعية بالرياض
Poison Control & Medical forensic Chemistry Center, Riyadh
CMIA is non-competitive sandwich assay technology
(Pre-Tigger Solution) (Tigger Solution) Activiation
Al-nafisah, B. PCC, Riyadh(2015)

15. ARCHITECT ci4100 Made by Abbott Very big instrument
مركز مراقبة السموم والكيمياء الطبية الشرعية بالرياض
Poison Control & Medical forensic Chemistry Center, Riyadh
ARCHITECT ci4100
Made by Abbott
Very big instrument
Measures multiple therapeutic drugs quantitatively
Also measures major drugs of abuse DOA (Sensitivity problem for some tests)
Very fast
Take many unlimited number of samples (good)
Requires external water supply !!!
Requires complex daily maintenance
Complex software (Requires careful and long training process)
Chemiluminescent microparticle immunoassay (CMIA) is the main technique for most TDM measurements
Al-nafisah, B. PCC, Riyadh(2015)

16. Others Immunoassays RIA: Radioimmunometric assays
مركز مراقبة السموم والكيمياء الطبية الشرعية بالرياض
Poison Control & Medical forensic Chemistry Center, Riyadh
Others Immunoassays
RIA: Radioimmunometric assays
Not commonly used any longer due to waste disposal issues
use radioactivity to detect the presence of the analyte.
A gamma counter is then used to measure the amount of radioactivity in the sample as counts per minute (CPM).
QC and standards must be performed with each run, leading to extra labor and cost.
ACMIA: Affinity Chrome-Mediated Immunoassay.
ACMIA is a technique to measure drug concentrations in which free and drug-bound antibody enzyme conjugates are separated using magnetic (chrome) particles.
Al-nafisah, B. PCC, Riyadh(2015)

17. PETINIA: Particle Enhanced Turbidimetric Inhibition Immunoassay
مركز مراقبة السموم والكيمياء الطبية الشرعية بالرياض
Poison Control & Medical forensic Chemistry Center, Riyadh
PETINIA: Particle Enhanced Turbidimetric Inhibition
Immunoassay
This method uses the creation of light scattering particles to measure drug levels. The latex particle-bound drug binds to the drug-specific antibody, forming insoluble light-scattering aggregates. This causes an increase in the turbidity of the reaction mixture.
CEDIA: Cloned enzyme donor immunoassay
(CEDIA) methodology is a novel approach which uses the DNA technology to produce homogenous enzyme immunoassays for drugs
Al-nafisah, B. PCC, Riyadh(2015)

18. The Biochip The Biochip is the foundation of Biochip Array Technology.
مركز مراقبة السموم والكيمياء الطبية الشرعية بالرياض
Poison Control & Medical forensic Chemistry Center, Riyadh
The Biochip
The Biochip is the foundation of Biochip Array Technology.
A single 9x9mm biochip acts as the reaction vessel, replacing multiple cuvettes.
Randox biochips are pre-fabricated with an array of discrete test regions (DTR’s)
Randox biochips currently hold up to 25 tests - 23 tests and 2 internal controls.
One biochip is used per patient sample.
Biochip manufacturing processes are accredited to UKAS and ISO13485.
Al-nafisah, B. PCC, Riyadh(2015)

19. Evidence Biochip Carrier
مركز مراقبة السموم والكيمياء الطبية الشرعية بالرياض
Poison Control & Medical forensic Chemistry Center, Riyadh
Evidence Biochip Carrier
Al-nafisah, B. PCC, Riyadh(2015)

20. مركز مراقبة السموم والكيمياء الطبية الشرعية بالرياض
Poison Control & Medical forensic Chemistry Center, Riyadh
Al-nafisah, B. PCC, Riyadh(2015)

21. مركز مراقبة السموم والكيمياء الطبية الشرعية بالرياض
Poison Control & Medical forensic Chemistry Center, Riyadh
DRUG DETCTION PERIODS
Al-nafisah, B. PCC, Riyadh(2015)

22. مركز مراقبة السموم والكيمياء الطبية الشرعية بالرياض
Poison Control & Medical forensic Chemistry Center, Riyadh
Sampling
Type of Samples received:
CRUDES, BIOLOGICALS and SEIZED TOOLS
Samples comes from hospitals forensic medicine administration and from different government sectors such as:
-Ministry of Interior ( General Directorate of Narcotics Control
and Police offices )
-Customs (Sea & Land ports)
-Other Government sectors
Al-nafisah, B. PCC, Riyadh(2015)

23. CRUDE SAMPLES - Khat Sampling Specifications:
مركز مراقبة السموم والكيمياء الطبية الشرعية بالرياض
Poison Control & Medical forensic Chemistry Center, Riyadh
CRUDE SAMPLES
Received from different sectors
Samples of seized crude materials such:
- Khat
- Cannabis
- Captagon tablets
- Alcohols
- Any seized materials and tools etc..
Sampling Specifications:
Sealed and secure
Privacy (no personal data)
Enough (Fixed amount for each drug)
e.g 5 grams khat
Comes with a letter from requesting sector
Received only from AUTHORIZED person
Al-nafisah, B. PCC, Riyadh(2015)

24. مركز مراقبة السموم والكيمياء الطبية الشرعية بالرياض
Poison Control & Medical forensic Chemistry Center, Riyadh
BIOLOGICAL SAMPLES
Include body fluids (i.e. blood, urine, gastric lavage, etc.) and tissue samples (Biopsies and Autopsies)
- Hospitals samples:
TDM , (ET) , Employees screening and police suspected cases
- Forensic medicine administration samples
Al-nafisah, B. PCC, Riyadh(2015)

25. BIOLOGICAL SAMPLES (FORENSIC)
مركز مراقبة السموم والكيمياء الطبية الشرعية بالرياض
Poison Control & Medical forensic Chemistry Center, Riyadh
BIOLOGICAL SAMPLES (FORENSIC)
Purpose: Employee screening , police suspected cases, forensic medicine (FM).
Urine sample is the main sample for Narcotics analysis
Blood sample is the main sample for alcohol analysis
For FM, all body fluids (urine, blood, Vitreous humor) and any other body parts or excretions can be received. NOTE: Crude samples can be received in some occasions to help in the detection of the death cause !!
Al-nafisah, B. PCC, Riyadh(2015)

26. BIOLOGICAL SAMPLES (FORENSIC) cont.
مركز مراقبة السموم والكيمياء الطبية الشرعية بالرياض
Poison Control & Medical forensic Chemistry Center, Riyadh
BIOLOGICAL SAMPLES (FORENSIC) cont.
Sampling specifications for employee screening:
2 urine samples
Request form
Labeled correctly
Biohazard bag and cold container
Authorized person
Sampling specifications for police cases:
2 urine samples for narcotics and 2 blood samples for alcohol
All papers completed (request from police, request from hospital, sample collection witness form)
Finger prints of the suspect on the samples and request
No delay
Al-nafisah, B. PCC, Riyadh(2015)

27. BIOLOGICAL SAMPLES (FORENSIC) cont.
مركز مراقبة السموم والكيمياء الطبية الشرعية بالرياض
Poison Control & Medical forensic Chemistry Center, Riyadh
BIOLOGICAL SAMPLES (FORENSIC) cont.
Sampling specifications for FM:
Any human or crude samples can be received
Request form completed by forensic doctor (dead person information, description of the body during sampling, any related information)
Labeled correctly
No delay
Biohazard container for tissue transportation and cold container
Authorized person
Al-nafisah, B. PCC, Riyadh(2015)

28. Right dose (in case of Emergency Toxicology & TDM) Right route
مركز مراقبة السموم والكيمياء الطبية الشرعية بالرياض
Poison Control & Medical forensic Chemistry Center, Riyadh
Five “Rights”
Right Sample
Right drug
Right dose (in case of Emergency Toxicology & TDM)
Right route
Right time
Al-nafisah, B. PCC, Riyadh(2015)

29. مركز مراقبة السموم والكيمياء الطبية الشرعية بالرياض
Poison Control & Medical forensic Chemistry Center, Riyadh
Al-nafisah, B. PCC, Riyadh(2015)

30. Conclusion Immunoassay excluded negative samples.
مركز مراقبة السموم والكيمياء الطبية الشرعية بالرياض
Poison Control & Medical forensic Chemistry Center, Riyadh
Conclusion
Immunoassay excluded negative samples.
Immunoassay evolves continuously.
Result obtained within short-time (minute).
Easy to use.
No require for extraction procedure.
No hazard chemicals used.
Al-nafisah, B. PCC, Riyadh(2015)

31. THANK YOU مركز مراقبة السموم والكيمياء الطبية الشرعية بالرياض
Poison Control & Medical forensic Chemistry Center, Riyadh
THANK YOU
Al-nafisah, B. PCC, Riyadh(2015)