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2017年1月5日 星期四

[醫如往常] Senior Clerkship Day 108: Pathology: Kidney

林醫師
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Sinus fat
Papilla pelvis
Pyramid

Gerota's fascia
T12-L3
Perirenal, pararenal fat

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Endothelial fenestrae 70-100nm
Mesangial

GBM
Lamina rara interna
Lamina densa
Lamina rara externa

Podocytes foot process 20-30nm visceral epithelium
Parietal epithelium
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Macula densa of distal tubule chemoreceptor
JGA baroreceptor
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90% renal cortex composes tubules
Proximal tubule, brush border 比較紅

Shock segments
Toxin continuous
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Interstitium
Ccr, BUN
Per min
1000cc renal artery
900cc renal vein
GFR 100
Urine 1
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Renal artery
Seg
Interlobar
Arcuate
Intralobar
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Percutaneous biopsy

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HE
Silver
Masson Trichrome
PAS
3um

Immunofluorescence
IgG, IgM, IgA, C3, C1q, kappa, lambda
Tubules deposition nonspecific
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Subepi.  Membrane
Mesengial IgA
Subendo lupus, MPGN

DIf
Cryo
Acetone
PBS
FITC
PBS
PermaFluor
Dark

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Goodpasture disease
Goodpasture syndrome (GPS; also known as Goodpasture’s disease, antiglomerular basement antibody disease, or anti-GBM disease) is a rare autoimmune disease in which antibodies attack the basement membrane in lungs and kidneys, leading to bleeding from the lungs and kidney failure. It is thought to attack the alpha-3 subunit of type IV collagen, which has therefore been referred to as Goodpasture's antigen.[1] Goodpasture syndrome may quickly result in permanent lung and kidney damage, often leading to death. It is treated with immunosuppressant drugs such as corticosteroids and cyclophosphamide, and with plasmapheresis, in which the antibodies are removed from the blood.
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EM
Glutaraldehyde
1um, 50-70nm for EM
Thin basement disease
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RPGN Ccr兩倍 in  3 month. Crescent
Type 1
Goodpasture
Linear deposition of IgG in GBM
NC1 domain

Type 2
Immune complex
SLE, IgA, type MPGN

Type 3
Pauci immune

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MPGN
Type 2 DDD, dense deposit disease
 Lamina densa, Bowman's capsule, renal tubules
Recurrence after transportation

C3 glomerular deposition atypical HUS
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Thin basement disease
400 200 nm
Type 4 collagen
Alpha 3,4 subunit defect
Deafness

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Alport syndrome
X linked
Alpha 5 subunit

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15% AR type
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DM nephropathy


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Myeloma
Kappa 多
Light chain
Bence Jones protein
Cast nephropathy

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Amyloid
Beta birefringence
AA, AL type most common
Inflammation AA 未開發
Light chain AL 已開發 lambda

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ATIN, AIN
Chinese herb nephropathy

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Granular kidney
Hypertension
Compensatory hypertrophy

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Rejection
器官經濟學
Endothelial double contour
C4d
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RCC 70%
Clear cell 70%

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UC

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[醫如往常] Senior Clerkship Day 107: Pathology: Mixed Germ cell tumor

    SPC 梁醫師
    Mixed Germ cell tumor 40M left retroperitoneal tumor with flank pain
    AFP4000 ng/ml
    Dopamine 550 ug/24h
    Rule out
    1. Castleman disease
    2. Psoas fibrous tumor
    3. Leiomyosarcomasarcoma
    4. Liposarcoma

    Calcification
    Vessel invasion
    ------------
    Mature teratoma GFAP
    Immature
    Microcystic
    Schiller-Dual Body (glomerular like)
    CD30 embryonal carcinoma
    Yolk sac tumor cd117
    Choriocarcinoma HSD3D1
    ------------
    ------------
    年紀越大, yolk sac tumor ratio越大
    Adult may combine hematopoietic neoplasms
    ------------
    Pediatric type
    Age>12 is adverse
    I(12p) is rare
    Sacrococcygeal>>mediastinum
    If it combines YST or other type, not a bad sign
    Conservative approach
    ------------
    Children prepubertal type
    6 year survival rate >80% with platinum
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    Extragonal adult type
    比照non seminoma處理
    常常是occult testes
    ------------
    EC 是不好的
    ------------
    Puberty 也可能長prepubertal type
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    Germ cell neoplasm in situ 是post pubertal 特徵
    ------------
    1cm3, 送cytogenetics
    20000自費
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[醫如往常] Senior Clerkship Day 107-108: Pathology: Breast

    Morning Meeting
    LAM; Lymphangioleiomyomatosis
    ------------
    Paraganglioma
    GATA 3 +
    S100 +
    Ck -
    ------------
    Breast
    Diabetes mastopathy
    Lymphocyte infiltration
    Big fibroblast
    ------------
    Parotids sebaceous lymphadenoma
    ------------
    Skin hyperpigmentation
    Fine chromatin
    Tdt trace
    CD3 -, 79a -
    CD123, 56, 4 +
    Blast
    Plasmacytoid dendritic cell neoplasm
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    Haematoma
    Cartilage 旁邊有東西
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    連醫師
    Breast cancer
    MRM
    Simple
    Partial/ BCT, lumpectomy
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    Facial margin
    ------------
    Serial slicing
    Med. to lat.
    3-5mm
    May skip
    ------------
    RCC Stage 1<7cm
    ------------
    Breast Mass 10cm, may be DCIS
    Clinical, pathology discrepancy

    外面醫院考慮成本 只能切五片
    NTUH切四十片
    ------------
    Random for multifocal
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    ALND
    至少要10顆
    小於十顆會被核刪
    Multislice 2mm
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    美國病理最便宜的180美金
    台灣從一千到五千點
    ------------
    2003開始sentinel lymph node. 通常2,3顆, 最多做過10顆
    Blue cassette
    500 um

    可能false negative tumor塞滿

    IHC 染ck

    Isolated tumor cell clusters <200 cells, <0.2mm
    pN0 i+
    ITCs
    可是NEJM2009 還是有差異
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    Van Nuys Prognostic classification
    RT for DCIS
    Age
    Grade
    Margin
    Size
    ------------
    ER, PR
    Her2
    Ki67 proliferation
    14% low
    20% high
    ------------

    FISH 可以與二十年前蠟塊
    PCR
    Microarray fresh tissue RNA
    ------------
    UCLA Jews Her2 tyrosine kinase
    Amplification, overexpression
    Chromosome 17
    Her2/centromere>2.0
    Counts 20 cells
    一萬元

    IHC 2+ 做FISH
    3+ 直接用Herceptin 80萬
    (Protein unstable)

    Doxorubicin(Adriamycin)
    Herceptin Trastuzumab
    都有cardiotoxicity

    Heart development Her2
    ------------
    Imatinib (Glivec) CML

    ------------
    Colon
    EGFR 60%
    Ras 40%
    Raf

    ------------
    Lung
    EGFR exon 18,19,20,21
    19 5 deletion
    21 1 deletion l858r

    T790m
    ------------
    Quality control sensitivity
    ------------
    Ewing sarcoma
    T 22, 11
    PCR
    FISH
    ------------

    ------------
    Day 108
    Morning Meeting
    Breast cancer with SPC; Solitary papillary carcinoma

    Increased, in situ, borderline, invasive
    • CAIX
    • MCT4
    Decreased

    ------------
    Invasive tumor with SPC >
    Pure in situ Spc with conventional DCIS

    • Mucinous type
    • WT1, Muc2

    CA9 loss of basement membrane (type iv collagen)
    • Old
    • Spindled tumor
    • ER, PR,chromogranino

    • Smaller size <20mm
    • Lower grade grade 1
    • Older age
    ------------
    Type B neuroendocrine
    • Synaptophysin
    • Chromogranin A
    • CD56
    • NSE
    ------------
    No significant difference between
    Invasive carcinoma with SPC and without SPC
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    Prostate幾乎都有neuroendocrine change
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[醫如往常] Senior Clerkship Day 106: Pathology: markers

孫法醫
Frozen
O.C.T:optimal cutting temperature compound -19--21
Molecule Liquid Nitrogen -85
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Special stain
Congo red: Amyloid
Copper: Wilsons' disease
PAS; Periodic acic Schiff stain: glycogen, MPS, capsule, fungus
DPAS; diastase : salivary
Silver: hepatoma
Orcein: elastic fiber, aneurysm, AVM
Von Kossa 照光: Calcium
Warthin-Starry: spirochette
PTAH(Phosphotungstic 鎢 acid haematoxylin): striation (rhabdomyosarcoma), fibrin (DIC)

Fresh tissue:
Fat stain
Thio-acetylcholine esterase (AchE): megacolon, lamina propria
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IHC
CD30: Anaplastic large cell lymphoma (ALCL)
classical Hodgkin Lymphoma Reed-Sternberg cells

CD45: CLA; common leukocyte antigen
CD68: macrophage
CD117(c-kit): GIST

plasmacytoma lambda常見

CK7: UGI
CK20: LGI
Ki-67: proliferation
P63: breast myoepithelial cells
GFAP: glioma
ALK: Lung
P40: SCC
MSH6: MSI; microsatellite instability
TTF-1: Thyroid, lung adenocarcinoma
Synaptophysin, Chromogranin: Neuroendocrine peptides
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Positive, negative control
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白色 biopsy
大件檢體
綠色 切兩片
藍色 牙科
粉紅色 autopsy
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10% neutral formalin (4% formaldehyde) 10x tissue + phosphatate buffer
滲透 1mm/ 1 hr
極限:五小時 上下加起來1cm
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Pancreas autolysis, fat necrosis
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Alcohol dehydration
Large cell -> small cell
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兩小時內
Bouin's solution-> bone marrow, spermatozoa
Masson's trichrome refixation

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Carnoy's solution -> EBV -> NPC
Acetic acid
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吳法醫師
CD 79a pan B cells
CD 20 mature B cells
CD10 follicle cells
CD23 dendritic cells,  marginal type small BCL
BCL-2 apoptosis

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Myocarditis治療
Hypothermia
ECMO Bypass



[醫如往常] Senior Clerkship Day 105: Lab Medicine: 國家級臨床試驗研究中心

國家級臨床試驗研究中心

FFPE Formalin-fixed, paraffin-embedded
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一次八千元
EGFR
2573
2369
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Kras +egfr 16000
四天發報告
四年現在兩萬例

NGS 一次十萬
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助理教授
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CR complete
PR partial
SD stable
PD; Progress disease
Response rate cr+pr
Control sd pr cr

Tki 一顆2000
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IVD In vitro diagnosis
LDT lab developed test
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Mass diagnosis

DNA Sample
Amplification
Detection, probe在突變前一個nucleotide
Analysis
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Kras smoking

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Cancer panal 52
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Clinical interpretation based report of NGS

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Afatinib 楊志新成名
10-12 month

T790m  mutation 60%
Azd9291
12-23 weeks

C797s mutation
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Drug resistance

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Circulation tumor dna; ct DNA
Cell free dna ; cf Dna
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Liquid biopsy

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PD1 只對20%有用
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[醫如往常] Senior Clerkship Day 103: Lab Medicine: Antibiotic sensitivity test

CLSI AST Breakpoints
Antibiotic sensitivity test
 劉醫師
Class ABC 藥物
O 研究藥物
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Disk dilution method 只會給大小, 不知道MIC
Broth dilution method
E test 用濾紙
Vitek
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BAP Agar
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MIC minimal inhibitory concentration
  • MIC50
  • MIC90
  • Mic epidemiological distribution, cutoff value (wild, mutant )
MBC minimal bactericidal concentration
MPC mutation prevention concentration
Serum bactericidal titer , for infective endocarditis
  • 一週沒改善,就要開刀去除biofilm
  • 現在很少做SPT
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Epidemiology
Pharmaceutical consideration
  • Time>MIC越久越好
  • Cmax/MIC
Clinical evaluation
Post market surveillance PMS
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Inhibitory quotient

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Post antibiotics effect

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Clinical success
USA 90%
ROC 80%

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Correlation

Oxacilin correlation很差,要用cefazolin做disk dilution
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現在有些標誌會改變42
但機器只能做到<4

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Intracellular bacteria做出來有效, 實際還是無效

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不能穿過BBB

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鄭醫師
Maryland general hospital

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雪梨宣言
Statement of death

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ISO 15189
Lab ethics
http://www.taftw.org.tw/wSite/ct?xItem=334&ctNode=177&mp=1 
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每分鐘20管

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H. Pylori test
紅外線200萬
質譜儀700萬
成本六百元
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False positive
Urease producing bacteria
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False negative
PPI
Gastic surgery
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Occult blood test
Chemical test10元 GI
  • Vit C false negative


Immunochemistry 70元 low GI
健保50點
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CA 72.4 gastric cancer, gastritis
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HCC, F18沒有用
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CA19-9
HbA1C升高也會高

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邵醫師
Galactomannan GM
Aspergillosis

Blood
Broncho lavage BAL 最敏感
CSF

Detect GAF tetramer

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CGD, chronic granumatous disease

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False positive
Antibiotics,
Other mycosis
比菲德氏菌

False negative

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Plasma
Serum
BAL
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Population 決定ppv,npv
常跟論文結果相反 , 因為族群不同,論文大多是探討screening
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Elisa

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Fungus PCR去年上市
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[醫如往常] Senior Clerkship Day 102: Lab Medicine: 細菌室

細菌室 薛P
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CSF 絕對不能冰,一小時送到檢醫部
  • Pneumococcus
  • Meningococcus
  • HiB

  • E coli
  • Listeria
  • KP
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CSF半小時送給細菌室 馬上接種
Gram stain危急值半小時內醫師要確認
沒確認打電話叫臨床看病歷系統
CAP要求99.9%
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Urine要冰,也是一小時送到
Loop 0.001ml
>10 0000 CFU
不能驗gonococcus(因為冰了會死掉),要用discharge culture, chocolate medium

一般Urine不做厭氧菌,做了也是汙染菌(unless suprapubic aspiration)
Vaginal discharge也不做厭氧菌
Stool做厭氧則是為了Clostridium difficile
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NTUH每天220culture,一天40個陽性,一半白天,一半晚上
60%來台大前就用過抗生素,要用resin血瓶

左右手各一套2-4瓶,陰性菌,陽性菌,黴菌,TB

Blood culture 也是一小時
Time to positive TTP
KP TTP七小時內的死亡率是TTP七小時以上的兩倍

Gram stain 30分鐘確認
Error 3%

導管菌血症TTP兩小時positive是導管感染

正常80%一天長, 95%兩天長
  • 細菌五天發no growth
  • TB 42天發no growth (最快7天長出來)
  • 黴菌28天發no growth (fungus都是需氧)
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Sputum要冰
所以養不出pneumococcus, HiB
找到yeast通常沒有意義,不會發報告
sputum不做厭氧菌,因為口腔一堆厭氧菌
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The wenstein–Jensen medium, more commonly known as LJ medium, is a growth medium[1] specially used for culture of Mycobacterium species, notably Mycobacterium tuberculosis.
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Q score
Documentation: 
Bartlett developed the Q score to grade the quality of a specimen sent for culture based on the Gram stain appearance. This can help determine if the specimen is good, fair or poor.
Specimen selection: A specimen from the lower respiratory tract or a superficial wound.
Gram stain examination: 10 to 20 low power (10x objective) microscopic fields
Parameters:
(1) average number of neutrophils present per low power field
(2) average number of squamous cells present per low power field
(3) presence of ciliated respiratory epithelial cells if a sputum specimen
(4) leukopenia due to disease or therapy
Parameter
Finding
Points
average number of neutrophils
0 (none)
0

1 to 9 (few)
+1

10 - 24 (moderate numbers)
+2

>= 25 (many, numerous)
+3
average number of squamous cells
0 (none)
0

1 to 9 (few)
-1

10 - 24 (moderate numbers)
-2

>= 25 (many, numerous)
-3
composite Q score =
= (points for average number of neutrophils) + (points for average number of squamous cells)
Interpretation:
• minimum score: -3
• maximum score: +3
• The higher the score the better the specimen.
A specimen with a composite score >= +1 should be cultured.
• A sputum specimen from a leukopenic patient with ciliated respiratory epithelial cells should be cultured.
• A composite Q score that is 0 or negative is probably a superficial sample that may not be a reliable specimen and so is usually not cultured.


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培養結果的判讀沒有所謂的黃金標準,常用的判別方式大致依病人的臨床狀況、分離菌種及陽性培養套數做判斷[1,3,6,7,9]。常見陽性的指標如下:1.病人具有敗血症症狀如發燒、體溫過低、白血球過多或過少、低血壓。2.分離出的菌種不像污染菌,如Staphylococcus aureus、Candida spp.、Streptococcus pneumoniae、Escherichia coli、其他腸內菌科及Pseudomonas aeruginosa 90%以上為致病菌;Strep. agalactiae、Strep. pyogenes、Listeria monocytogenes、Nesseria meningitidis、Nesseria gonorrhoeae、Haemophilus influenzae、Bacteroides fragilis group、Cryptococcus neoformans則皆為致病菌幾乎不可能為汙染菌[6]。3.由不同抽血部位抽取的檢體,同時分離出相同的病 原菌。但如為白血病患者則上述臨床症狀則可能不適用。

CoNS、Corynebacterium spp.、Propionibacterium acnes及碳疽桿菌之外的Bacillus spp.、Micrococcus spp.、viridans streptococci及Clostridium perfringens[6]為污染的可能性較高,其中又以C. spp.、P.acnes、M. spp.及碳疽桿菌之外的Bacillus spp.的致病機率最低。但上述各菌種為真正致病菌的百分比,在每個研究的結論並不一致,所以醫師無法僅以菌種名稱決定分離菌株的臨床意義。
只以分離套數來區分CoNS是否為污染菌,結果並不正確,依據文獻結果:兩套培養皆分離CoNS和兩套培養中只有1套分離的污染可能性,分別為31%及87%[4];12-31%的污染個案為多套分離,而12-35%的真正感染菌只有從單套培養分離[3,4,7]。故只能說愈多套分離其為污染菌的可能性愈小。而若只有抽單套培養又培養陽性時,其污染的可能性則為67-89%,判讀其臨床意義的困難度更高,故不建議只送單套血液培養[4,7]。另外以分子生物方法,分析同一病人由導管處及週邊血管同時分離的S. epidermidis時,發現4/14為不同型;另一研究則發現插管後14天,55%至少兩套分離的CoNS為不同型,可能為污染或多重菌株感染[7],故僅以分離套數來界定分離菌株的臨床意義確實有其缺點存在。

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AST; antimicrobial susceptibility test: 一種抗生素Etest一隻$150,健保只給付220
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Bruker Maldi-Tof MS
一台800萬,做一次的成本2
但沒有買機器的話,做一次200

美國沒有的Burkholderia pseudomallei
Vibrio vulnificus
Penicillin marafi (AIDS常見)
質譜儀容易跑錯
------------

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黃醫師
FUO
murmur, cordae tenden rupture
IE,

注意水質,legionella

Campylobacter 兩個彎
Helicobacter 多個彎

Candin不能cryptococcus
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Catheter infection
TTP 早兩小時
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PJP PCR
成本600
1000
長庚4500
Dyspnea spo2 90%
CXR 不明顯 clear

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TSI Mueller-Hinton medium
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Urine 看到candida很正常
看到CryptococcusDissemination

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