ASCP Recalls as of August’ 2013 1.How is Calcium measured in ISE? Ionized Calcium 2.Know the picture of: E. coli:
E. nana:
E. histolytica:
3.a!imum inter"al which a recipient sample may#e used for crossmatchin$ if the patient has #een recently transfused% has #een pre$nant? 3 days &. 'efri$eration 'efri$eration of #lood samples what is a(ected?
inc potassium, dec glucose , dec enzymes tryglicerides tryglicerid es not aected
).'$t strip *+,% Ictotest *-,% hat does this mean?
! "ierence in sensiti#ity le#els ! Rgt strip can detect detect 0$% mg &dl of 'iliru'in 'iliru'in ()ile Ictotest from from 0$0% !0$1 mg& dl$ ! Ictotest Ictotest is a ta'let test 'ased 'ased on diazotizat diazotization* ion* +- 'lue 'lue or purple purple color in .0 seconds
/.hat is a(ected in lactic acidosis?
! a form of meta'olic acidosis ! dec P/, inc actate, dec ygen ! deep and rapid 'reat)ing , #omiting, and a'dominal pain ! caused 'y dia'etic etoacidosis, etoacidosis, li#er or idney diseases, medication + p)enformin-, /I4 drugs + antiretro#irals- , arsenic poisoning 0.rocedure for actic cid test ! used as an indirect assessment of oygen le#el in tissues and to determine t)e cause of lactic acidosis 4. Culture media for e$ionella - 'uered c)arcoal!yeast etract +5C67- agar media 5. C6 #lood was drawn 7 18 m% pooled 7 11:38 am. atient has !ray 7 2 pm% what will you do with the product? ! transfuse t)e 'lood 'efore ray 18. En9ymes:
estroy ; 8 9 S "uy Enhance ; R) e(is I :idd
Cold Anti'odies; e(is S
8
9
R/ :ell
Pi
I
"uy :idd
11.
! 7 ! R= ! I8 ! Infectious /epatitis ! eprosy ! 8alaria ! Pregnancy ! Aging process ! Pneumococcal pneumonia 12. 'ead a#out C'
13. 'ead a#out 'I = radioimmunoassay
1&. Study > test attern with picture:
=eed'ac 6his is an e!ample of a mi! of )omogeneous and specled A9A patterns$
In this sample notice the specled > is the dominant pattern in the interphase cells *a, and some speclin$ in the area outside of the chromosomal area of the mitotics *#,. lso notice the smooth stainin$ of the chromosomal area of the metaphase mitotic cells *c,. 6his represents the presence of a homo$eneous > pattern.
a. Perip)eral +rim- = the central protein of the nucleus is only sli$htly stained or not stained at all % #ut nuclear mar$ins @uoresce stron$ly and appear to e!tend into the cytoplasm + i.e. anti = > ! associated with S7 in the acti"e sta$e of the dis. and in S>ogren’s dis$ #.
/omogenous + diuse- = the whole nuicleus @uoresces e"enly + i.e. anti = > anti = Histone anti = >
+ + +
typically seen in 'heumatoid disorders Inc. titres are su$$esti"e of SE ec. titres may#e found in SE% '% SAo$renBs syndrome and i!ed Connecti"e 6issue is. * C6,
+
c. Specled + a $rainy pattern with numerous round dots of nuclear @uorescence% without stainin$ of the nucleoli + i.e. anti = '> anti = Sm . SpeciDc for SE + occurs in the presence of anti#ody to any e!tracta#le nuclear anti$en de"oid of > or histone + the anti#ody is detected a$ainst the saline e!tracta#le nuclear anti$ens + antio#odies to Sm anti$ens is hi$hly speciDc for SE and as a marer anti#ody.
+
+ d. 9ucleolar + a few round% smooth nucleoli that "ary in si9e will @uoresce when e!amined with FG. + i.e. anti+ nucleolar + present in )8 with Scleroderma% SAo$renBs syndrome% SE
+
+
e. Anti centromere + discrete and specled + Hi$hly selecti"e for C'ES6 +
rimary illiary Cirrhosis
In order to understand the ANA test (antinuclear antibody test), it is first important to understand different types of antibodies. •
nti#odies are proteins% produced #y white #lood cells% which normally circulate in
•
the #lood to defend a$ainst forei$n in"aders such as #acteria% "iruses% and to!ins. utoanti#odies% instead of actin$ a$ainst forei$n in"aders% attac the #odyJs own
•
cells. 6his is an a#normality. ntinuclear anti#odies are a uniue $roup of autoanti#odies that ha"e the a#ility to attac structures in the nucleus of cells. 6he nucleus of a cell contains $enetic material referred to as > *deo!yri#onucleic acid,.
An ANA test (antinuclear antibody test) can be performed on a patient's blood sample as part of the diagnostic process for certain autoimmune diseases. How the Test Is erformed To perform the ANA test, sometimes called !ANA (fluorescent antinuclear antibody test), a blood sample is drawn from the patient and sent to the lab for testing. "erum from the patient's blood specimen is added to microscope slides which ha#e commercially prepared cells on the slide surface. If the patient's serum contains antinuclear antibodies, they bind to the cells (specifically the nuclei of the cells) on the slide. A second antibody, commercially tagged with a fluorescent dye, is added to the mi$ of patient's serum and commercially prepared cells on the slide. The second (fluorescent) antibody attaches to the serum antibodies and cells which ha#e bound together. %hen the
slide is #iewed under an ultra#iolet microscope, antinuclear antibodies appear as fluorescent cells. •
If @uorescent cells are o#ser"ed% the > test is considered positi"e.
•
If @uorescent cells are not o#ser"ed% the > test is considered ne$ati"e.
ANA Titer A titer is determined by repeating the positi#e test with serial dilutions until the test yields a negati#e result. The last dilution which yields a positi#e result (fluorescence obser#ed under the microscope) is the titer which gets reported. Here is an e$ample& & positi#e & positi#e &* positi#e &+ positi#e & positi#e (reported titer) &- negati#e arts of an ANA eport An ANA report has three parts& •
ositi"e or ne$ati"e
•
If positi"e% a titer is determined and reported
•
6he pattern of @uorescence is reported
"ignificance of ANA attern ANA titers and patterns can #ary between laboratory testing sites, perhaps because of #ariation in methodology used. These are the commonly recogni/ed patterns& •
/omogeneous + total nuclear @uorescence due to anti#ody directed a$ainst
•
nucleoprotein. Common in systemic lupus erthematosus *SE,. Perip)eral + @uorescence occurs at ed$es of nucleus in a sha$$y appearance. nti+
•
> anti#odies cause this pattern. lso common in SE. Specled + results from anti#ody directed a$ainst di(erent nuclear anti$ens.
•
9ucleolar + results from anti#ody directed a$ainst a speciDc '> conD$uration of the nucleolus or anti#ody speciDc for proteins necessary for maturation of nucleolar '>. Seen in patients with systemic sclerosis *scleroderma,.
ositi#e ANA Test esult 0$plained
Antinuclear antibodies are found in patients who ha#e #arious autoimmune diseases, but not only in autoimmune diseases. Antinuclear antibodies can be found also in patients with infections, cancer, lun$ diseases, gastrointestinal diseases, hormonal diseases, blood diseases, sin diseases, and in elderly people or people with a family history of rheumatic disease. Antinuclear antibodies are actually found in about 12 of the normal population, too. ANA test results are 3ust one factor considered when a diagnosis is being formulated. A patient's clinical symptoms and other diagnostic tests must also be considered by the doctor. 4edical history is also significant because some prescription drugs can cause 5drug6 induced antinuclear antibodies5. Incidence of ANA in 7arious 8iseases "tatistically6spea9ing, the incidence of positi#e ANA test results (in percent per condition) is& •
Systemic lupus erythematosus *lupus or SE, + o"er 5)
•
ro$ressi"e systemic sclerosis *scleroderma, + /8+58
•
'heumatoid arthritis + 2)+38
•
SAo$renJs syndrome + &8+08
•
•
Lu"enile arthritis + 1)+38
"ubsets of the ANA tests are sometimes used to determine the specific autoimmune disease. !or this purpose, a doctor may order anti6ds8NA, anti6"m, "3ogren's syndrome antigens(""A, "":), "cl6; antibodies, anti6centromere, anti6histone, and anti6N. The ANA test is comple$, but the results (positi#e or negati#e, titer, pattern) and possible subset test results can gi#e physicians #aluable diagnostic information.
Several different serum tests are used to detect autoimmunity. These are conditions where the immune system acts directly against the bodys own tissues. One test, the ANA, or anti-nuclear antibody test, detects antibodies that are directed against various components of the nucleus of the cell. These include antibodies that have been formed against double-stranded or singlestranded DNA two ways in which the cells DNA can be found in the serum after being released from old and dying cells!. Other components of the nucleus such as histones are also released from old cells and can also become targets of the immune response. "hen they appear they may be mar#ers for e$cess or inappropriate immune responses directed against ones own tissues. %hysicians in our group test for various autoantibodies in order to characteri&e patients as those who might have a tendency for autoimmune responses. Those who test positive have been found to have a higher ris# for recurrent pregnancy loss and are more li#ely to benefit from therapeutic interventions see diagram!.
Consequences
-Antinuclear Antibody ANA! positive, spec#led pattern. -Autoantibody to DNA leads to inflammation in the placenta. -Autoimmune disease screening in the woman is negative No evidence of lupus or rheumatoid arthritis!. A blood test determines the presence of antibodies to polynucleotides, histones and DNA. This process involves running '( different tests on a sample of blood. The presence of antibodies is also tested for by doing the ANA test. This is a less sensitive test but one that many doctors have already done on their patients before we ever see them. The test is reported as a titer and a pattern. Any titer above )*+ is significant. The titers can get into the thousands such as )*',. This simply means that the test is positive when the blood serum is diluted many times. The pattern is reported as homogeneous, nucleolar or spec#led* -omogeneous* the antibody is to the ss DNA or ds DNA. -Nucleolar* the antibody is directed to the polynucleotides. -Spec#led* the antibody is directed against the histones. Some women demonstrate a mi$ed pattern of spec#led/homogeneous. These same antibodies appear positive in women with lupus, rheumatoid arthritis, 0rohn1s disease and other autoimmune diseases. They are usually in high titers. %regnancy losses, infertility and 234 failures cause the titers to be much lower and a low positive titer does not mean that you have or are getting an autoimmune disease5 however, this is ruled out during the testing. 2n women with autoimmune diseases these antibodies cause inflammation in 6oints and organs. 2n women with no autoimmune diseases but a positive antibody, the antibody causes inflammation around the embryo at the time of implantation or in the placenta after implantation. This inflammation is e$actly the same as occurs if you get a splinter under your fingernail. The tissue around the splinter gets hot, red and swollen and it happens 7uic#ly.
1). 'ead a#out IC for suscepti#ility test to report in micro#iolo$y
1/. Epstein arr Girus% How to detect? Know the procedure.
! infectious mononucleosis ! test to detect; a- 8ono spot '- C5C c- 754 serology ! can )elp detect if an indi#idual )as an infection due to 754, and if t)ey are prone to future infections due to dormant #irus$ ! 4CA!Ig8 4A!Ig? @ests !!!!!!!!!! )elp to identify current infection 7A!" 759A @est !!!!!!!!!!!!!! )elp to d future infection due to an eisting dormant #irus$ d- t)roat culture e- i#er prole !
10.H$1c )% #lood $luscose 288 $mMdl. hat does this mean?
! ?lucose normal for t)e past mont), currently )ig)$ 14. Know how to answer acid M$as case study.
:no( t)e 9ormal 4alue to 'e a'le to ans(er case study$ P/ B $3% ! $D% PC2 B 3% ! D% /C3 B 22 ! 2. 15. 18 sodium hypochlorite = for cleaning surfaces 28. Know how to identify pictures of euemias C C 21. Identify N discrepancy case study ! ()at to do net
!()at is causing t)e discrepancy 22. Identify Entero#acteria or$anism.: Ese t)e Fas) card
7:7
7SCP
:7S
7$ coli :7S S)igella Salmonella 7d(ardsiella
I ! !
P8P
P8P:
8 !
C; ! ! !
4 ! ! !
S:6
67SC
:77PS
23. Identify picture of microor$anism. ?ram +- or gram +!2&.
iliru#in re hepatic Hepatic ost Hepatic
inc
Fro#ilino$en
dec
2). E(ect of e!posure of #lood to air ! C02 dec, PC2 dec, P/ inc
! dec Ca, dec acid p)osp)atase, dec glucose, dec 'iliru'in
2/. 10 etosteoid is produced in ! Adrenal gland 20. 'ead on eil
! !
test for Ricettsia!!!!!! @yp)us fe#er t)is test in#ol#es testing of certain strains of Proteus #ulgaris
24. 'ead on Syphilis ! @reponema pallidum ! Reagin ! non treponemal anti'odies ! "areld 8icroscopy ! test of c)oice for patient (it) S6 ! 9on @reponemal 8et)od; ! a Focculation + or agglutination test 14"R ! can 'e used to test for CS= 2RPR ! contains c)arcoal * can’t 'e used for CS= * Causes of =alse +! 7
! R= ! I8 ! Infectious /epatitis ! eprosy ! 8alaria ! Pregnancy ! Aging process ! Pneumococcal pneumonia 25. Steps in a$$lutination
!
Sensitization ; 1st step in a$$lutination ; physical attachments of anti#ody molecules to anti$ens on the 'C mem#ranes
+
attice
38. ilution: 1st tu#e /th tu#e 1.1 Serum 8.5 diluent
2nd tu#e
3rd tu#e
1;320
31. hat causes syno"ial @uid tur#idity? Choices: a,
! !
)th tu#e
8.) serum 8.) diluent
hat is the dilution in the / th tu#e?
! ! !
&th tu#e
c-
S= !!! Increase content of )yaluronic acid + mucin8ucin clot test !!! precipitation of S= (it) (ea acetic acid Immunoglo'ulins , immune complees, complement !!!!!! produced 'y cells in t)e imFamed >oints 9ormal S= does not clot , #iscous, and clear S= t)at clots suggests t)e presence of syno#itis and is cause 'y 'rinogen
32. Oreen top tu#e % #lood is collected and refri$erated for 3 hours. Should you not accept? y answer: plasma s)ould 'e separated 'efore refrigeration
33. ala#soption test?
=ecal fat
3&.
! /aemo)ilus + / $ parainFuenza, ap)rop)ilus, parap)rop)ilus ! Actino'acillus + actinomycetemcomitans ! Cardio'acterium )ominis ! 7ienelle corrodens ! :ingella + :$ ingae3/. re"ents replication of "irus? Interferon 30. 'esult of hemodilution? ! lo( serum electrolytes 34. s a result of hemolysis + !
freezing canm cause )emolysis dec :, dec "/, dec AS@, normal uric acid
34. automaterd method for measurin$ Chloride which $enerates sil"er ions in the reaction. + cystic 'rosis + Coulometry 35. urpose of documentation lo$. &8. ia$nosis of a case study: ?lomerulonep)ritis Pyelonep)ritis Cystitis &1.
=lo( cytometry is the most relia#le method of those listed. It is a uantitati"e method% whereas Keihauer+ete and the rosette test are "ery su#Aecti"e tests.
&3. /emoglo'in electrop)oresis uses an electric Deld to separate hemo$lo#in molecules #ased on di(erences in net electrical char$e. 6he rate of electrophoretic mi$ration is also dependent on the ionic radius of the molecule% the "iscosity of the solution throu$h which it is mi$ratin$% the electrical Deld stren$th% temperature% and the type of supportin$ medium used. &&. @)in!layer c)romatograp)y is particularly useful as a tool in the identiDcation of: + ru$s &)$ :no( 5lood Panel &/. Know aternity 6estin$ &0. Study ea in lood anin$ &4. achines set 7 1)8 wa"elen$ht% then wa"elen$th 7 8 when used. hat is the pro#lem?
&5. 'elease of results to whom? y answer : entist )8. 'ead on immunoassay
)1. lactam 6est ++++++
$rm *+,
)2. Study nion Oap
)3.6roponin = en9yme in I ele"ated the lon$est )&. Study I en9ymes
)). i"er isease : study other y answer: 6 S6 )/. Study
nemia of chronic diseases 6halassemia
)0. onoclonal $raph . hat to do ne!t? + multiple myeloma + presence of ence Aones protein in the urine + monoclonal $ammopathy
8one marrow cytology in a dog with multiple myeloma. There are large numbers of plasma cells 9! in the aspirate, some of which are binucleate. ematopoietic cells both myeloid :;< and erythroid :=
. 4yeloma
with roulea$ cells
Labs: Findings
A. "erum rotein 0lectrophoresis and =rine rotein electrophoresis for 4onoclonal ea9 . 4 rotein in either serum or urine& >;2 of patients . "erum 4 rotein by electophoresis (+2) or immunofi$ation (>-2) -. =rine 4 rotein by electrophoresis& ;12 :.
:. :one 4arrow Aspiration and biopsy <. eripheral "mear 4yeloma
"uggested seCuence of immunologic testing & electrophoresis
4 spi9e on serum protein
Serum:
+Immunoelectrophoresis +ImmunoD!ation +Puantitation of immuno$lo#ulins #y radial immunodi(usion or nephelometry + Screenin$ for cro$lo#ulins + etermination of serum "iscosity of I$% I$% or I$O % or si$ns and symptoms su$$esti"e of Hyper"iscosity
Urine: 6 "creening of urine for increased protein, e.g. sulfosalicylic acid 6 Total protein assay of a * hour urine specimen 6 =rinary protein electrophorsis 6 =rinary immunoelectrophoresis 6 Immunofi$ation
)4. Salmonella in the diarrhea. Know how to I.
)5. ro"idencia or$anism
/8. roteus Gul$aris or$anism
/1. Culture media for e$ionella?
/3. Hematolo$y: 6ar$et Cells IC C S *-, and SF> *-, is what? Smud$e cells i$ latelet
/&.
Inc. ES' causes: Study othe causes y answer Q Inc altelet
/). 'ouleau! formation due to ? Choose: a, Slow or fast smearin$ #, n$le of the smear c, d,
//. ormal 6IC
/0. Hereditary ChromatosisQ C242R H/3 S/)C
++++++++ in@ammatory process
> analysis of the followin$
/4. Know the cases for eferral of #lood donors: y answer:
t with CG
/5. 6ests a(ected #y #lood hemolysis? + Increase; K% % S6% lasma H$# ! "ecrease ; 6& + =alsely 7le#ated: hosphorus% 6otal protein% l#umin% a$nesium% Calcium% cid hosphatase 08. (ected #y e!posure of li$hts to #lood: + "ecrease : Gitamin and /% eta+carotene% horphyrins% iliru#in Should #e wrapped in aluminum foil immediately after collection.