csf by asif
TRANSCRIPT
![Page 1: Csf by asif](https://reader035.vdocuments.site/reader035/viewer/2022062304/55ce56d1bb61ebc3468b4649/html5/thumbnails/1.jpg)
CSF Examination
Muhammad Asif Zeb lecturer Hematology IPMS-KMU
![Page 2: Csf by asif](https://reader035.vdocuments.site/reader035/viewer/2022062304/55ce56d1bb61ebc3468b4649/html5/thumbnails/2.jpg)
ContentsIntroductionFunction Sample CollectionCSF Lab ExaminationChemical AnalysisSerological ExaminationOther TechniquesReferences
![Page 3: Csf by asif](https://reader035.vdocuments.site/reader035/viewer/2022062304/55ce56d1bb61ebc3468b4649/html5/thumbnails/3.jpg)
GENERAL INFORMATION:
Both the brain and spinal cord are covered by three protective membranes referred to as the meninges. The outermost layer is called the dura mater and is composed of tough connective tissue. The middle layer is the arachnoid named for it spider web like appearance. The delicate innermost layer which is in direct contact with the brain and spinal cord is called the pia mater. An inflammation of the meninges is referred to as meningitis.
![Page 4: Csf by asif](https://reader035.vdocuments.site/reader035/viewer/2022062304/55ce56d1bb61ebc3468b4649/html5/thumbnails/4.jpg)
Between the arachnoid layer and the pia mater is a space called the subarachnoid space. It contains a clear, colorless fluid referred to as Cerebrospinal Fluid (CSF). CSF is produced in the ventricles of the brain by a collection of rich vascular protrusions called the choroid plexus. Excess CSF is continuously reabsorbed by arachnoid villi and returned to the venous system thus maintaining a consistent amount of fluid
![Page 5: Csf by asif](https://reader035.vdocuments.site/reader035/viewer/2022062304/55ce56d1bb61ebc3468b4649/html5/thumbnails/5.jpg)
FUNCTIONS OF CSFTHE TWO CHIEF FUNCTIONS OF CSF (SPINAL) FLUID ARE:1. To protect and cushion the brain and the spinal cord against possible injury.
2. To deliver nutrients from the blood to the Central Nervous System and removeswastes
![Page 6: Csf by asif](https://reader035.vdocuments.site/reader035/viewer/2022062304/55ce56d1bb61ebc3468b4649/html5/thumbnails/6.jpg)
Normal CSFThin, colourless, clear fluid
CSF is the 3rd major fluid of the bodyAdult volume 90-150 mLNeonate volume 10-60 mL
0-5 WBC’s /mm3 (neonates 0-30/ mm3 ) (Lymphocytes & monocytes)
Occasional ependymal or choroid plexus cellsProtein 15-45mg/dlGlucose 50-80mg/dlChloride 113-130 mEq/LSterile
![Page 7: Csf by asif](https://reader035.vdocuments.site/reader035/viewer/2022062304/55ce56d1bb61ebc3468b4649/html5/thumbnails/7.jpg)
Lumber puncture or collection of CSf
The procedure used to obtain cerebrospinal fluid is referred to as Lumbar Puncture or a Spinal Tap.
![Page 8: Csf by asif](https://reader035.vdocuments.site/reader035/viewer/2022062304/55ce56d1bb61ebc3468b4649/html5/thumbnails/8.jpg)
PurposeThe purpose of CSF analysis is to diagnose medical Disorders that affect the CNS.
Viral and Bacterial Infections.Tumors of Nervous System.Syphilis, a Sexually transmitted disease.Bleeding around the brain and spinal cord.Multiple sclerosis,a disease that affects the myelin coating of the nerve fibers.Guilain-Barre syndrome,an inflammation of the nerves.
![Page 9: Csf by asif](https://reader035.vdocuments.site/reader035/viewer/2022062304/55ce56d1bb61ebc3468b4649/html5/thumbnails/9.jpg)
Collection Technique
![Page 10: Csf by asif](https://reader035.vdocuments.site/reader035/viewer/2022062304/55ce56d1bb61ebc3468b4649/html5/thumbnails/10.jpg)
full flexion
![Page 11: Csf by asif](https://reader035.vdocuments.site/reader035/viewer/2022062304/55ce56d1bb61ebc3468b4649/html5/thumbnails/11.jpg)
![Page 12: Csf by asif](https://reader035.vdocuments.site/reader035/viewer/2022062304/55ce56d1bb61ebc3468b4649/html5/thumbnails/12.jpg)
Level of entry
![Page 13: Csf by asif](https://reader035.vdocuments.site/reader035/viewer/2022062304/55ce56d1bb61ebc3468b4649/html5/thumbnails/13.jpg)
Spinal needle
![Page 14: Csf by asif](https://reader035.vdocuments.site/reader035/viewer/2022062304/55ce56d1bb61ebc3468b4649/html5/thumbnails/14.jpg)
Spinal needle
![Page 15: Csf by asif](https://reader035.vdocuments.site/reader035/viewer/2022062304/55ce56d1bb61ebc3468b4649/html5/thumbnails/15.jpg)
Under strict aseptic precautions.,
![Page 16: Csf by asif](https://reader035.vdocuments.site/reader035/viewer/2022062304/55ce56d1bb61ebc3468b4649/html5/thumbnails/16.jpg)
![Page 17: Csf by asif](https://reader035.vdocuments.site/reader035/viewer/2022062304/55ce56d1bb61ebc3468b4649/html5/thumbnails/17.jpg)
![Page 18: Csf by asif](https://reader035.vdocuments.site/reader035/viewer/2022062304/55ce56d1bb61ebc3468b4649/html5/thumbnails/18.jpg)
Collecting the CSF
![Page 19: Csf by asif](https://reader035.vdocuments.site/reader035/viewer/2022062304/55ce56d1bb61ebc3468b4649/html5/thumbnails/19.jpg)
19
Collection: 3-10 ml is collected
Tube 1Tube 1 Chemistry & Chemistry & immunologyimmunology
Tube 2Tube 2 MicrobiologyMicrobiology
Tube 3Tube 3 Cytology and Cytology and HematologyHematology
![Page 20: Csf by asif](https://reader035.vdocuments.site/reader035/viewer/2022062304/55ce56d1bb61ebc3468b4649/html5/thumbnails/20.jpg)
AnalysisPhysical examinationChemical examinationMicroscopic examination
![Page 21: Csf by asif](https://reader035.vdocuments.site/reader035/viewer/2022062304/55ce56d1bb61ebc3468b4649/html5/thumbnails/21.jpg)
Appearance of CSFclear and colourless (normal); bright red - This indicates the presence of blood. The appearance of fresh blood in all three tubes supports the diagnosis of a subarachnoid hemorrhage. xanthochromic (yellow) - jaundice
turbid - This indicates the presence of white cells and is suggestive of a CNS infection.
![Page 22: Csf by asif](https://reader035.vdocuments.site/reader035/viewer/2022062304/55ce56d1bb61ebc3468b4649/html5/thumbnails/22.jpg)
Presence of clotNo clotIf present indicate high protein contentCondition With patient traumatic tapComplete spinal blockage Tuberculosis meningitisBacterial infection
![Page 23: Csf by asif](https://reader035.vdocuments.site/reader035/viewer/2022062304/55ce56d1bb61ebc3468b4649/html5/thumbnails/23.jpg)
Amount of CSFHow much collected
![Page 24: Csf by asif](https://reader035.vdocuments.site/reader035/viewer/2022062304/55ce56d1bb61ebc3468b4649/html5/thumbnails/24.jpg)
Microscopic examinationCell counts
Noramal less than 5/ul TLC count RBC count Smear preparation for Differential count to see the morphology
of WBC,sUsually few lymphocytes are present
![Page 25: Csf by asif](https://reader035.vdocuments.site/reader035/viewer/2022062304/55ce56d1bb61ebc3468b4649/html5/thumbnails/25.jpg)
How to count TLC and RBC???
![Page 26: Csf by asif](https://reader035.vdocuments.site/reader035/viewer/2022062304/55ce56d1bb61ebc3468b4649/html5/thumbnails/26.jpg)
Clinical significance of Increase TLC
Neutrophili leukocytosis =Neisseria meningitis, Hemophilus influenza and E. coliLymphocytic/monocytic leukocytosis=T.b, Viral infection, fungus or with syphilisMalignancy= Leukemia,
![Page 27: Csf by asif](https://reader035.vdocuments.site/reader035/viewer/2022062304/55ce56d1bb61ebc3468b4649/html5/thumbnails/27.jpg)
Leukocyte-differential
Most patients with bacterial infections=neutrophils while fungal, tubercular and viral infection = lymphocytes.
![Page 28: Csf by asif](https://reader035.vdocuments.site/reader035/viewer/2022062304/55ce56d1bb61ebc3468b4649/html5/thumbnails/28.jpg)
Leukocyte-differential (Contd..)
other conditions.LeukemiaPlasma cell leukemiaPresence of tumor in brain.Metastasis to CSF
![Page 29: Csf by asif](https://reader035.vdocuments.site/reader035/viewer/2022062304/55ce56d1bb61ebc3468b4649/html5/thumbnails/29.jpg)
Gram stain and cultureA Gram’s stain should be perform on all CSF.Cultures are also set up in order to retrieve and identify organisms present in the CSF.
If the physician suspects a viral, mycobacterial, or rickettsia organism,
it is important that special culture techniques be followed for isolation of these organisms.
![Page 30: Csf by asif](https://reader035.vdocuments.site/reader035/viewer/2022062304/55ce56d1bb61ebc3468b4649/html5/thumbnails/30.jpg)
Chemical analysis
CSF PROTEIN DETERMINATIONCSF Glucose DETERMINATION
![Page 31: Csf by asif](https://reader035.vdocuments.site/reader035/viewer/2022062304/55ce56d1bb61ebc3468b4649/html5/thumbnails/31.jpg)
ProteinNormal CSF protein concentration is 15 - 45 mg/dl. It is derived from the plasma.
Increased levels are associated with:- infection- altered capillary permeability- decreased absorption- local biosynthesis of gamma globulin
![Page 32: Csf by asif](https://reader035.vdocuments.site/reader035/viewer/2022062304/55ce56d1bb61ebc3468b4649/html5/thumbnails/32.jpg)
Sulfosalicylic acid testTake 3 ml of 3% sulfosalicylic acid in a tube add 1 ml of supernatant clear CSF in it.Cloudiness indicate the presence of protein
![Page 33: Csf by asif](https://reader035.vdocuments.site/reader035/viewer/2022062304/55ce56d1bb61ebc3468b4649/html5/thumbnails/33.jpg)
Principle (Biuret Method): Any compound(proteins) containing three or more peptide bonds reacts with alkaline copper tartrate reagent to form a blue to purple coloured
substance. The intensity of colour produced is
proportional to the number of peptide bonds reacting and, therefore, to the amount of proteins.
![Page 34: Csf by asif](https://reader035.vdocuments.site/reader035/viewer/2022062304/55ce56d1bb61ebc3468b4649/html5/thumbnails/34.jpg)
Biuret MethodProcedure: To 2 ml CSF add 2 ml 10%trichloracetic acid, mix well and allow to stand for 5 min. Centrifuge at high speed and discard the supernatant. Mark this tube containing precipitate as test. Take another test tube and mark it blank. To both tubes add 1 ml 15% NaOH. Shake the “test” tube to dissolve them precipitate. Add 0.5 ml 5% Copper sulphate and 4 ml distilled water. Mix thoroughly and centrifuge at high speed. Transfer the supernatant to corresponding clean tubes marked. Read absorbance of the “test” against “blank” in a colorimeter at 550 nm.
![Page 35: Csf by asif](https://reader035.vdocuments.site/reader035/viewer/2022062304/55ce56d1bb61ebc3468b4649/html5/thumbnails/35.jpg)
CSF GLUCOSE DETERMINATION
• Normal CSF glucose is: 60 to 70 % of the patient's serum glucose concentration.
It is generally decreased in bacterial meningitis, as well as in meningitis associated with a fungus, tuberculosis or syphilis.
![Page 36: Csf by asif](https://reader035.vdocuments.site/reader035/viewer/2022062304/55ce56d1bb61ebc3468b4649/html5/thumbnails/36.jpg)
Chemical analysis (Contd..)
Decreased protein levels are less commonly encountered.Common reasons are removal of large amounts of CSF and leakage of CSF caused by trauma.Glucose concentrations can be determined using the same methodologies as used for blood.The most common finding is a low glucose concentration associated with most types of CNS infections.If patient is hyperglycemic, then the CSF glucose concentrations will like wise be elevated and an infection may be masked.If the patient is hypoglycemic the physician may be misled by a low CSF glucose.
![Page 37: Csf by asif](https://reader035.vdocuments.site/reader035/viewer/2022062304/55ce56d1bb61ebc3468b4649/html5/thumbnails/37.jpg)
Chemical analysis (Contd..)
Lactate determination is considered to be of some use by physicians in differentiating bacterial, tuberculous and fungal meningitis from viral meningitis.The lactate concentration in viral meningitis is generally normal or low in comparison to that in bacterial, tuberculous, or fungal meningitis.
![Page 38: Csf by asif](https://reader035.vdocuments.site/reader035/viewer/2022062304/55ce56d1bb61ebc3468b4649/html5/thumbnails/38.jpg)
![Page 39: Csf by asif](https://reader035.vdocuments.site/reader035/viewer/2022062304/55ce56d1bb61ebc3468b4649/html5/thumbnails/39.jpg)
Serologic examinationSerologic examination of CSF is typically involved with diagnosis of neurosyphilis.A fluorescent treponemalantibody absorption (FTA-ABS) test for CSF has also been developed.The VENERAL DISEASE RESEARCH LAB. (VDRL) Test is also used.TPHA test or TPPA test for syphlus
![Page 40: Csf by asif](https://reader035.vdocuments.site/reader035/viewer/2022062304/55ce56d1bb61ebc3468b4649/html5/thumbnails/40.jpg)
Other techniquesIt is important to recognize other techniques that have been used and may still be used for CSF.
Immunoelectrophorsis (CIE), latex agglutination tests for bacterial antigens, enzyme linked immunosorbent assay (ELISA) and limulus of gram-negative bacterial).
The nucleic acid probes using PCR are the newest of the techniques being investigated for indentification of specific infectious agents.
![Page 41: Csf by asif](https://reader035.vdocuments.site/reader035/viewer/2022062304/55ce56d1bb61ebc3468b4649/html5/thumbnails/41.jpg)
Thank You