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.Maximum interval which a recipient sample maybe used for crossmatching if the patient has been recently transfused, has been pregnant? 3 days

4. Refrigeration of blood samples what is affected?

inc potassium, dec glucose , dec enzymes

tryglicerides not affected

5.Rgt strip (-), Ictotest (+), What does this mean?

- Difference in sensitivity levels- Rgt strip can detect 0.5 mg /dl of bilirubin while Ictotest from 0.05 -0.1 mg/ dl. - Ictotest is a tablet test based on diazotization; (+) blue or purple color in 60 seconds

6.What is affected in lactic acidosis? - a form of metabolic acidosis- dec PH, inc Lactate, dec Oxygen- deep and rapid breathing , vomiting, and abdominal pain- caused by diabetic ketoacidosis, liver or kidney diseases, medication ( phenformin), HIV drugs ( antiretrovirals) , arsenic poisoning

7.Procedure for Lactic Acid test- used as an indirect assessment of oxygen level in tissues and to determine the cause of lactic acidosis

8. Culture media for Legionella- buffered charcoal-yeast extract (BCYE) agar media

9. CPT blood was drawn @ 10 Am, pooled @ 11:30 am. Patient has xray @ 2 pm, what will you do with the product?- transfuse the blood before xray

10. Enzymes: Destroy = M N S Duffy Enhance = Rh Lewis I Kidd

Cold Antibodies: M N Pi I Lewis S Warm Antibodies : RH Kell Duffy Kidd

11.False (+) for RPR ( Rapid Plasma Reagin ) Non treponemal test for Rickettsia

- LE- RF- IM- Infectious Hepatitis- Leprosy- Malaria- Pregnancy- Aging process- Pneumococcal pneumonia

12. Read about PCR

13. Read about RIA radioimmunoassay

14. Study ANA test Pattern with picture:

FeedbackThis is an example of a mix of homogeneous and speckled ANA patterns.

In this sample notice the speckled ANA is the dominant pattern in the interphase cells (a) and some speckling in the area outside of the chromosomal area of the mitotics (b).Also notice the smooth staining of the chromosomal area of the metaphase mitotic cells (c). This represents the presence of a homogeneous ANA pattern.

a. Peripheral (rim) the central protein of the nucleus is only slightly stained or not stained at all , but nuclear margins fluoresce strongly and appear to extend into the cytoplasm i.e. anti DNA associated with SLE in the active stage of the dis. and in Sjogrens dis.

b. Homogenous ( diffuse) the whole nuicleus fluoresces evenly i.e. anti DNA anti Histone anti DNP

typically seen in Rheumatoid disorders Inc. titres are suggestive of SLE Dec. titres maybe found in SLE, RA, Sjogrens syndrome and Mixed Connective Tissue Dis. ( MCTD)

c. Speckled - a grainy pattern with numerous round dots of nuclear fluorescence, without staining of the nucleoli i.e. anti RNP anti Sm . Specific for SLE occurs in the presence of antibody to any extractable nuclear antigen devoid of DNA or histone the antibody is detected against the saline extractable nuclear antigens antiobodies to Sm antigens is highly specific for SLE and as a marker antibody.

d. Nucleolar - a few round, smooth nucleoli that vary in size will fluoresce when examined with UV. i.e. anti- nucleolar present in 50% with Scleroderma, Sjogrens syndrome, SLE

e. Anti centromere - discrete and speckled Highly selective for CREST

Primary Billiary Cirrhosis

In order to understand the ANA test (antinuclear antibody test), it is first important to understand different types of antibodies. Antibodiesare proteins, produced by white blood cells, which normally circulate in the blood to defend against foreign invaders such as bacteria, viruses, and toxins. Autoantibodies, instead of acting against foreign invaders, attack the body's own cells. This is an abnormality. Antinuclear antibodiesare a unique group of autoantibodies that have the ability to attack structures in the nucleus of cells. The nucleus of a cell contains genetic material referred to as DNA (deoxyribonucleic 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 PerformedTo perform the ANA test, sometimes called FANA (fluorescent antinuclear antibody test), a blood sample is drawn from the patient and sent to the lab for testing. Serum from the patient's blood specimen is added to microscope slides which have 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 mix of patient's serum and commercially prepared cells on the slide. The second (fluorescent) antibody attaches to the serum antibodies and cells which have bound together. When the slide is viewed under an ultraviolet microscope, antinuclear antibodies appear as fluorescent cells. If fluorescent cells are observed, the ANA test is considered positive. If fluorescent cells are not observed, the ANA test is considered negative.ANA TiterA titer is determined by repeating the positive test with serial dilutions until the test yields a negative result. The last dilution which yields a positive result (fluorescence observed under the microscope) is the titer which gets reported. Here is an example:1:10 positive1:20 positive1:40 positive1:80 positive1:160 positive (reported titer)1:320 negativeParts of an ANA ReportAn ANA report has three parts: Positive or negative If positive, a titer is determined and reported The pattern of fluorescence is reportedSignificance of ANA PatternANA titers and patterns can vary between laboratory testing sites, perhaps because of variation in methodology used. These are the commonly recognized patterns: Homogeneous- total nuclear fluorescence due to antibody directed against nucleoprotein. Common in systemic lupus erthematosus (SLE). Peripheral- fluorescence occurs at edges of nucleus in a shaggy appearance. Anti-DNA antibodies cause this pattern. Also common in SLE. Speckled- results from antibody directed against different nuclear antigens. Nucleolar- results from antibody directed against a specific RNA configuration of the nucleolus or antibody specific for proteins necessary for maturation of nucleolar RNA. Seen in patients with systemic sclerosis (scleroderma).Positive ANA Test Result ExplainedAntinuclear antibodies are found in patients who have variousautoimmune diseases, but not only in autoimmune diseases. Antinuclear antibodies can be found also in patients with infections,cancer,lung diseases, gastrointestinal diseases, hormonal diseases, blood diseases,skin diseases, and in elderly people or people with a family history of rheumatic disease. Antinuclear antibodies are actually found in about 5% of the normal population, too.ANA test results are just 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. Medical history is also significant because some prescription drugs can cause "drug-induced antinuclear antibodies".Incidence of ANA in Various DiseasesStatistically-speaking, the incidence of positive ANA test results (in percent per condition) is: Systemic lupus erythematosus (lupus or SLE)- over 95% Progressive systemic sclerosis (scleroderma)- 60-90% Rheumatoid arthritis- 25-30% Sjogren's syndrome- 40-70% Felty's syndrome- 100% Juvenile arthritis- 15-30%Subsets of the ANA tests are sometimes used to determine the specific autoimmune disease. For this purpose, a doctor may order anti-dsDNA, anti-Sm, Sjogren's syndrome antigens(SSA, SSB), Scl-70 antibodies, anti-centromere, anti-histone, and anti-RN.The ANA test is complex, but the results (positive or negative, titer, pattern) and possible subset test results can give physicians valuable 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 single-stranded 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. When they appear they may be markers for excess or inappropriate immune responses directed against ones own tissues. Physicians in our group test for various autoantibodies in order to characterize patients as those who might have a tendency for autoimmune responses. Those who test positive have been found to have a higher risk for recurrent pregnancy loss and are more likely to benefit from therapeutic interventions(see diagram).

Consequences

-Antinuclear Antibody (ANA) positive, speckled 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 27 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 1:40 is significant. The titers can get into the thousands such as 1:2,500. This simply means that the test is positive when the blood serum is diluted many times.

The pattern is reported as homogeneous, nucleolar or speckled:

-Homogeneous: the antibody is to the ss DNA or ds DNA.-Nucleolar: the antibody is directed to the polynucleotides.-Speckled: the antibody is directed against the histones.

Some women demonstrate a mixed pattern of speckled/homogeneous. These same antibodies appear positive in women with lupus, rheumatoid arthritis, Crohn's disease and other autoimmune diseases. They are usually in high titers. Pregnancy losses, infertility and IVF failures cause the titers to be much lower and a low positive titer does not mean that you have or are getting an autoimmune disease; however, this is ruled out during the testing.

In women with autoimmune diseases these antibodies cause inflammation in joints and organs. In 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 exactly 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 quickly.15. Read about MIC for susceptibility test to report in microbiology

16. Epstein Barr Virus, How to detect? Know the procedure.

- infectious mononucleosis- test to detect: a) Mono spot b) CBC c) EBV serology - can help detect if an individual has an infection due to EBV, and if they are prone to future infections due to dormant virus. - VCA-IgM VA-IgG Tests ---------- help to identify current infection EA-D EBNA Test -------------- help to dx future infection due to an existing dormant virus.

d) throat culture e) Liver profile-

17.HgA1c 5, blood gluscose 200 gm/dl. What does this mean?

- Glucose normal for the past month, currently high.

18. Know how to answer acid /gas case study.

Know the Normal Value to be able to answer case study. PH = 7.35 - 7.45 PCO2 = 35 - 45 HCO3 = 22 - 26

19. 10% sodium hypochlorite for cleaning surfaces

20. Know how to identify pictures of Leukemias ALL AML CLL CML

21. Identify ABO discrepancy case study - what to do next -what is causing the discrepancy 22. Identify Enterobacteria organism.: Use the flash card

EKE ESCP KES PMP PMPK SKY YESC KEEPS

I M V C:E. coli + + - -KES - - + +Shigella + + + +Salmonella - + - -Edwardsiella + + - -

23. Identify picture of microorganism. Gram (+) or gram (-)

24. Bilirubin Urobilinogen Pre hepatic Hepatic Post Hepatic inc dec

25. Effect of exposure of blood to air - C02 dec, PCO2 dec, PH inc

- dec Ca, dec acid phosphatase, dec glucose, dec bilirubin

26. 17 ketosteoid is produced in - Adrenal gland

27. Read on Weil Felix Reaction

- test for Rickettsia------ Typhus fever- this test involves testing of certain strains of Proteus vulgaris

28. Read on Syphilis- Treponema pallidum- Reagin - non treponemal antibodies- Darkfield Microscopy - test of choice for patient with SY- Non Treponemal Method: - a flocculation ( or agglutination ) test 1) VDRL - can be used to test for CSF 2) RPR - contains charcoal ; cant be used for CSF ; Causes of False (+)- LE- RF- IM- Infectious Hepatitis- Leprosy- Malaria- Pregnancy- Aging process- Pneumococcal pneumonia

29. Steps in agglutination

Sensitization= 1st step in agglutination= physical attachments of antibody molecules to antigens on the RBC membranes

Lattice Formation= establishments of cross links between sensitized particles and antibodies resulting in aggregation (clumping), is a much slower process thant the sensitization phase

30. Dilution: 1st tube 2nd tube 3rd tube 4th tube 5th tube 6th tube0.1 Serum 0.5 serum 0.9 diluent 0.5 diluent

What is the dilution in the 6th tube? 1:320

31. What causes synovial fluid turbidity? Choices: a) Fibrinogen b) crystals c) immunoglobulins d) fibrinogen III

SF --- Increase content of hyaluronic acid ( mucin) Mucin clot test --- precipitation of SF with weak acetic acid Immunoglobulins , immune complexes, complement ------ produced by cells in the imflamed joints Normal SF does not clot , viscous, and clear SF that clots suggests the presence of synovitis and is cause by fibrinogen

32. Green top tube , blood is collected and refrigerated for 3 hours. Should you not accept? My answer: plasma should be separated before refrigeration

33. Malabsoption test? Fecal fat

34. Fungus picture? My answer penicillium

35.Group of organism for HACEK

- Haemohilus ( H . parainfluenza, aphrophilus, paraphrophilus ) - Actinobacillus ( actinomycetemcomitans ) - Cardiobacterium hominis - Eikenelle corrodens - Kingella ( K. kingae)

36. Prevents replication of virus? Interferon

37. Result of hemodilution? - low serum electrolytes

38. As a result of hemolysis

- freezing canm cause hemolysis - dec K, dec LDH, dec AST, normal uric acid

38. automaterd method for measuring Chloride which generates silver ions in the reaction. - cystic fibrosis - Coulometry

39. Purpose of documentation log.

40. Diagnosis of a case study: Glomerulonephritis Pyelonephritis Cystitis 41. Fetal lung maturity

42. Which of the following methods isMOST reliable for determining the appropriate dosage of Rh immune globulin togiveto an identified Rh immune globulin candidate after delivery?

Flow cytometry is the most reliable method of those listed. It is a quantitative method, whereas Keihauer-Betke and the rosette test are very subjective tests.

43. Hemoglobin electrophoresis uses an electric field to separate hemoglobin molecules based on differences in net electrical charge. The rate of electrophoretic migration is also dependent on the ionic radius of the molecule, the viscosity of the solution through which it is migrating, the electrical field strength, temperature, and the type of supporting medium used.44. Thin-layer chromatography is particularly useful as a tool in the identification of: - Drugs45. Know Blood Panel46. Know Paternity Testing47. Study Weak D in Blood Banking48. Machines set @ 150 wavelenght, then wavelength @ 0 when used. What is the problem?

49. Release of results to whom?My answer : Dentist

50. Read on immunoassay

51. B lactam Test ------ grm (-)

52. Study Anion Gap

53.Troponin enzyme in MI elevated the longest

54. Study MI enzymes

55. Liver Disease : study other My answer: ALT AST

56. Study Ferritin and transferring levels in : Hemochromatosis Iron deficiency anemia Anemia of chronic diseases Thalassemia

57. Monoclonal graph . What to do next?

- multiple myeloma - presence of Bence jones protein in the urine - monoclonal gammopathy

Bone marrow cytology in a dog with multiple myeloma. There are large numbers of plasma cells (*) in the aspirate, some of which are binucleate. Hematopoietic cells (both myeloid [M] and erythroid [E]) are found in normal numbers and maturation sequence.

1. Myeloma Cells with rouleax cellsI. Labs: FindingsA. Serum Protein ElectrophoresisandUrine Proteinelectrophoresis for Monoclonal PeakA. M Protein in either serum or urine: 97% of patientsA. Serum M Protein by electophoresis (82%) or immunofixation (93%)A. Urine M Protein by electrophoresis: 75%B. Chemistry panel withSerum CalciumB. Hypercalcemiaa. Serum Calcium>11 mg/dl (present in 13% of patients)Renal Insufficiencya. Serum Creatinine>2 mg/dl (present in 23% of patients)C. Complete Blood Countwith plateletsNormochromicNormocytic Anemiaa. Hemoglobin50 mm/hour in most cases (except bence-jones Myeloma)G. Serum ViscosityH. UrinalysisBence-Jones Protein2. Rouleaux ofRed Blood Cells

Suggested sequence of immunologic testing : M spike on serum protein electrophoresis Serum:-Immunoelectrophoresis-Immunofixation-Quantitation of immunoglobulins by radial immunodiffusion or nephelometry- Screening for croglobulins- Determination of serum viscosity of IgM, IgA, or IgG , or signs and symptoms suggestive of Hyperviscosity

Urine: - Screening of urine for increased protein, e.g. sulfosalicylic acid - Total protein assay of a 24 hour urine specimen - Urinary protein electrophorsis - Urinary immunoelectrophoresis - Immunofixation

58. Salmonella in the diarrhea. Know how to ID.

59. Providencia organism

60. Proteus Vulgaris organism

61. Culture media for Legionella?

63. Hematology: Target Cells

DIC

CLL

PAS (+) and SUDAN (+) is what?

Smudge cells

Big Platelet

64. Inc. ESR causes: Study othe causes My answer ; Inc Paltelet

65. Rouleaux formation due to ? Choose:a) Slow or fast smearingb) Angle of the smear c) d)

66. Ferritin :

Increase Ferritin , Normal TIBC -------- inflammatory process

67. Hereditary Chromatosis; DNA analysis of the following C282Y H63D S65C

68. Know the cases for Deferral of blood donors: My answer: Pt with CMV

69. Tests affected by blood hemolysis? - Increase: K, LD, AST, Plasma Hgb - Decrease : T4 - Falsely Elevated: Phosphorus, Total protein, Albumin, Magnesium, Calcium, Acid Phosphatase

70. Affected by exposure of lights to blood: - Decrease : Vitamin A and B6, Beta-carotene, Phorphyrins, Bilirubin Should be wrapped in aluminum foil immediately after collection.