Transcript 微生物及免疫相關課程:教學目標

Introduction to clinical
serology and
immunology
Outline

Seat assignment and knowing each other.
 Course overview
 網路教學
 http://moodle.ncku.edu.tw
葉才明(Trai-Ming Yeh)教授
實驗室電話: 5778
 辦公室地點: 醫技系 館4樓5794室
 電子信箱 [email protected]
 學歷
1990美國德州大學微生物免疫學博士
1984台大微生物所寄生蟲組碩士
1980台大醫技系學士

How Was Clinical Laboratory Started?
Some pioneers foresee the need to bring
basic science into clinical medicine to help
attack clinical problems.
 Medical Technology: the bridge between
basic and clinical science

微生物及免疫相關課程:教學目標

微生物感染及免疫是醫學不可或缺的一環，
微生物學及臨床微生物學和臨床血清免疫
學與臨床病毒學更是在醫檢師專技高考六
科中的兩科。因此我們希望學生修完這些
相關課程後，對醫學微生物及免疫學有通
盤的認識，使之不但對於各項臨床微生物
檢驗（包括細菌、寄生蟲、病毒和黴菌）
及血清免疫檢驗的原理、技術及應用，可
以有相當程度的瞭解，並且能夠培養出對
此領域在工作及研究上的興趣及能力。
微生物及免疫相關課程特色

師資設備一流完善
 本課程參與老師均分別從事微生物及免疫相關
基礎及臨床工作相關設備完善，學生學習成果
良好。

授課實驗相輔相成
 學生實際動手從事各樣相關實驗，包括細菌、
寄生蟲、病毒和黴菌的各種培養或檢驗。

課程內容完整一貫
 含括基礎及臨床各種微生物及免疫學
課程規劃與架構

學生必須先修畢大一的基本科目如生物、
解剖、生理等，進而在大二下及大三上修
習基礎微生物學、寄生蟲學及免疫學，大
三下則將所學的基礎課程擴展為臨床微生
物學、臨床血清免疫學及臨床病毒學並配
合相關實驗課程。大四上則以實習為主，
實際在醫院學習各樣臨床微生物檢驗及血
清免疫檢驗。
相關課程
PROGRAM GOAL

To maintain a curriculum which will
provide graduates with the information of
basic principles and methods of
immunology and their clinical application
(for the board exam)
References
Textbook
Immunology and Serology in Laboratory
Medicine, Mary L. Turgeon, 4th ed., 2009
References
 Immunobiology, C. Janeway, 7th ed., 2008.
 Kuby Immunology, R. A. Coldsby, 6th ed. 2007.
 Medical Immunology, T. G. Parslw, D. P. Stites,
A.I. Terr, and J. B. Imboden 10th ed. 2001.
GRADING
Midterm Examination
40%
 Final Examination
40%
 Performance (attendance, home work and
participation)
20%

Definition of clinical serology

Literally, the study of serum. Refers to the study of
antigen-antibody responses which is important in clinical
medicine, especially in
(1) Diagnosis of infectious diseases by detecting
antibody against specific pathogen in serum.
(2) Monitor progress of disease and treatment
(3) Transfusion
(4) Health examination

Clinical immunology: The study of the immune function
and diseases of the immune system in an individual
Overview of immune system
Tissues and organs of the
immune system

Primary lymphoid
organs
 Thymus
 Bone

marrow
Secondary lymphoid
organ
 Spleen
 Lymph
nodes
General review of immune
response
Non-specific immune response (innate
immunity) (macrophage, dendritic cells,
neutrophils)
 Specific immune response (adaptive or
acquired immunity) (T cells and B cells)

 Memory
 Specificity
 Recognition
of "non-self"
Mechanisms of the non-specific
immune response

Humoral response
 Acute
phase proteins
 Complement

Cellular response
 Polymorphonuclear
neutrophils (PMN)
 Eosinophils
 mast
cells
 Basophils
 Mononuclear Phagocyte System
(Reticuloendothelial system)
Mechanisms of the specific
immune response
團結力量大
(Innate and adaptive immunity)
 Innate
immune response (macrophage,
dendritic cells, neutrophils)
 Pathogen associated molecule pattern
(PAMP)
 Pattern recognition receptor (PRR)
 Adaptive immune response (T cells and B cells)
 DNA rearrangement to generate T and B
cells receptor (TCR, BCR)
 Positive and negative selection
Innate immunity is required for the
induction of adaptive immunity
Kinetic of primary and secondary
antibody response
Immunity （免疫力）
was originally used to indicate exemption from
taxes and this meaning still exits in the term
"diplomatic immunity".
1. Innate (先天）vs Adaptive （後天或應變）
immunity
2. Humoral (體液） vs. Cellular (細胞）
immunity
3. Passive (被動） vs. Active (主動）
immunity

Safety and basic
techniques in the
immunology-serology
laboratory
General Technique of Serology

Sample: most are serum (no hemolysis, no
lipemia) , sometimes CSF or other body fluids.
Plasma contains fibrinogen which may form fibrin
in the presence of calcium and cause false
agglutination.
 Serum separation: blood clot 1 h at R.T.,
loosening the clot, centrifuge for 10 min at 9000xg
 Complement inactivation: complement may cause
prozone effect and leads to false negative. It can
however, be prevented by heating at 56C, 30 min
or adding choline chloride.
What is the “titer”?
目前對Ab的測定多採semiquantitative,通常用”titer(效價)”
表示
”產生陽性反應的最高稀釋倍數
(Serial dilution )”
What is the “titer”?

臨床檢驗
定性
(qualitative)
半定量
(semiquantitative)
Y/N , P/N
Ex.血型檢查
P/N,粗略表達
Ex.沉澱線
其含量,通常用視
(Ag-Ab)
覺判斷
定量
量的多寡
(quantitative)
Ex.glucose
Paired sera and sera conversion
Paired sera (two phase examination, acute
phase and convalescent phase) are required.
 Significant rise or drop in the titer (4-fold or
greater) indicates recent infection or vaccination.
 Constant titer indicates infection or vaccination
at an undetermined time.
 Seroconversion: antibody from negative to
positive.

Detection of antigen specific IgM
IgM is usually produced only early in
infection, would eliminate the need for a
second specimen. However, rheumatoid
factor (RF) may cause false positive and
excess antigen specific IgG may cause
false negative.
 RF is mostly IgM against self IgG

Screening vs. Confirmation test

Screening test
 high
sensitivity but low specificity
 Easy and cheaper for large scale test

Confirmation test
 Detect
more specific antigen and antibody
Sensitivity vs. specificity
TP: true positive
TN: true negative
FP: false positive
FN: false negative
Universal blood and body fluid
precautions
All human blood and other body fluids are
treated as potentially infectious for HIV,
HBV, HCV…etc (nosocomial transmission).
 HBV stable in dried blood up to 7 days in
RT, HIV 3 days and 1 week in aqueous
environment at RT.

Protective techniques for infectious
control
Glove
 Lab coats
 Facial barrier (mask)
 Hand washing: rub for at least 15 sec.
 Decontamination: 1:10 diluted household
bleach (0.5%NaOCl) kill HBV in 10 min
and HIV in 2 min.
 Needle precaution

Prophylaxis, medical follow-up, and
records of accidental exposure
Vaccination against HBV
 Hepatitis B immune globulin (HBIG) given
if accidental infection of HBV happen and
has no immunity against HBV.
 Free of charge HBV, HCV titers follow-up
 Injury report must be filed after parental
exposure.

Hazardous material and waste
management
Infectious waste: marked “Biohazard”,
autoclave
 Chemical hazards: material safety data
sheet (MSD), centralized store.
 Radioactive waste

Standard operation protocol (SOP)
A few things to discuss

Summer Biotechnology practice
course