Tests for Secondary Hemostasis PDF

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Institute of Health Technology, Dhaka

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blood coagulation hematology coagulation tests

Summary

This document details procedures and principles for a variety of blood coagulation tests. It includes the tests for the intrinsic and common pathways, plasma recalcification time, activated partial thromboplastin time, and various other tests. The document is written for an undergraduate-level class.

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Te sts f o r S eco n dary H em osta sis 0 8 TES T S F O R T H E I N TR IN SIC A N D C O M MON P A TH W AY1LE E A N D W HIT E W HO LE B LO OD C O AG ULA TIO N T IM E is b ase d o n t h e f a c t t h at w hen v e n ous b lo od i s p ut i n to a g la ss t u b e, i t w ill f o rm a s o...

Te sts f o r S eco n dary H em osta sis 0 8 TES T S F O R T H E I N TR IN SIC A N D C O M MON P A TH W AY1LE E A N D W HIT E W HO LE B LO OD C O AG ULA TIO N T IM E is b ase d o n t h e f a c t t h at w hen v e n ous b lo od i s p ut i n to a g la ss t u b e, i t w ill f o rm a s o lid c lo t Th e t im e r e q uir e d f o r t h is r e sp on se i s a m easu re o f t h e o ve ra ll i n trin sic a n d c o m mon p ath w ays I n se n sit iv e t o f a c to r d efic ie n cie s P ro ce d ure : W hole b lo od i s d is p en se d i n to t h re e 1 3 x10 0 m m o r 1 2 x75 m m t u b es ( t u b e 3 , t u b e 2 , t u b e In cu b ate i n a w ate rb ath a t 3 7°C f o r 5 m in ute s C heckin g i s d on e e ve ry 3 0 s e co n d s u n til a s o lid c lo t h as f o rm ed ( t u b e 1 , t u b e 2 , t u b e 3 ) N .V . 7 -15 m in ute s 2P IA SM A R EC ALC IF IC ATIO N T IM E A m od if ic atio n o f L -W c lo ttin g t im e Th is t e st i s b ase d o n t h e f a c t t h at e xc e p t f o r c alc iu m , P R P c o n ta in s a ll t h e c o m pon en ts n ece ssa ry f o r c re atin g a f ib rin c lo t Th e t im e r e q uir e d f o r b lo od t o c lo t a ft e r C a2 i s a d ded i s a g en era l m easu re o f t h e i n trin sic a n d c o m mon p ath w ay R efe re n ce r a n g es: P R P: 1 0 0-15 0 s e co n d s PPP: 1 3 0 -2 40 s e co n d s PR s h ould c lo t a t l e ast 2 0 s e co n d s f a ste r t h an P PP 3(A C TIV A TED ) P A RTIA L T H RO M BO PLA ST IN T IM E ___________________: a re l ip op ro te in s w hic h c an b e f r a c tio n ate d i n to l ip id s a n d p ro te in s Part ia l t h ro m bop la stin - o n ly t h e _ __ _________________ c o m pon en t i s p re se n t It s c re en s f o r f a c to r d efic ie n cie s i n t h e i n trin sic a n d c o m mon p ath w ays ( m easu re s a ll f a c to rs e xc e p t V II a n d X III) R eag en t c o n sis ts o f t w o c o m pon en ts P la te le t s u b stit u te ( p hosp holip id ) Activ a to rs : _ _______________ , _ _______________ , _ _______________ , _ _____________ __ , P ro ce d ure : P PP+ A PT r e ag en t ( p hosp holip id +a c tiv a to r+ C aC l2 ) N .V . 2 0 -4 5 s e co n d s U se d f o r m on it o rin g : _ ___________________________ TES T S F O R T H E E X TR IN SIC A N D C O M MON P A TH W AY1PR O TH RO M BIN T IM E Te sts f o r f a c to r I , I l, V , V ll a n d X d efic ie n cy ( E xtrin sic a n d C om mon ) Te st o f c h oic e f o r m on it o rin g a n tic o ag ula n t t h era p y w it h V it . K a n ta g on is t ( w arfa rin ) Pro ce d ure : P PP + P T r e ag en t ( T h ro m bop la stin +C aC l2 ) N .V . 1 0 -12 s e co n d s In te rn atio n al N orm aliz e d R atio P ro vid es a m ean s o f s ta n d ard iz in g P T r e p ort in g w orld w id e; n ot d ep en d en t o n Th ro m bop la stin r e ag en t U se d t o m on it o r c o um arin / w arfa rin t h era p y Th era p eu tic r a n g e i s d ep en d en t o n c o n d it io n b ein g t re ate d , b ut i s g en era lly c o n sid ere d to b e b etw een 2 .0 -3 .0 F o rm ula = ( P atie n t P T/ c o n tro l P T) I S I In te rn atio n al S en sit iv it y I n d ex ( IS I) f o r t h e t h ro m bop la stin r e ag en t i s p ro vid ed b y t h e m an ufa c tu re r a n d i s b ase d o n t h e W HO s ta n d ard s Th e c lo se r t h e I S I i s t o 1 , t h e m ore s e n sit iv e t h e r e ag en t O TH ER C O AG ULA TIO N T ES T S1ST Y PV EN T IM E Utiliz e s t h e p ow erfu l c o ag ula n t p ro p ert ie s o f R usse l's v ip er v e n om o b ta in ed f r o m t h e s n ake V ip era r u sse lli P R IN C IP LE : T h e v e n om i s c ap ab le o f b yp assin g t h e a c tio n o f f a c to r V II a n d d ir e ctly a c tiv a te s Fa c to r X t o X a It w as u se d t o h elp d is tin g uis h b etw een f a c to r X a n d V II d efic ie n cie s N .V . 6 -10 s e co n d s 25M U REA S O LU BIL IT Y T ES T A KA _ _____________________________ C lo ts f o rm ed I n n orm al p la sm a a re i n so lu b le i n 5 M u re a b ecau se o f t h e a c tio n f a c to r X III 3T H RO M BIN T IM E Use d t o s c re en f o r f ib rin og en d efic ie n cy a n d t h e p re se n ce o f f ib rin d eg ra d atio n p ro d ucts A ddit io n o f t h ro m bin t o p la sm a b yp asse s a ll c o ag ula tio n r e ac tio n s e xc e p t p oly m eriz a tio n o f fib rin og en D ysfib rin og en em ia , h yp ofib rin og en em ia , F D Ps, i m mun olo g ic a n tit h ro m bin s, a n d a b norm al g lo b ulin s w ill p ro lo n g t h ro m bin t im e PR O CED URE: P PP+ T h ro m bin r e ag en t N .V . 1 0 -14 s e co n d s 4REP TIL A SE T IM E Use d t o a sse ss f ib rin og en d efic ie n cy i n p atie n ts u n d erg oin g h ep arin t h era p y Reag en t: B oth ro p sa tro x s n ake v e n om Prin cip le : T h e v e n om h as t h ro m bin l i k e a c tiv it y a n d i s u n affe cte d b y h ep arin N .V . 1 0 -15 s e co n d s TH RO M BIN T IM EREP TIL A SE T IM EHep arin T h era p y Pro lo n g ed N orm al Fib rin S p lit P ro d ucts P ro lo n g edP ro lo n g ed H yp ofib rin og en em iaG re atly P ro lo n g ed P ro lo n g ed D ysfib rin og en em ia P ro lo n g ed G re atly P ro lo n g ed Im mun olo g ic a n tit h ro m bin s Pro lo n g ed N orm al 5C LA U S F I A SSA Y m od if ic atio n o f t h e T C T, r e co m men d ed p ro ce d ure f o r e stim atin g t h e f u n ctio n al f ib rin og en le ve l S A M PLE : P PP R EA G EN T: b ovin e t h ro m bin T h ro m bin r e ag en t c o n ce n tra tio n = 5 0 N IH u n it s /m L PPP t o b e t e ste d i s d ilu te d 1 :1 0 w it h O w en b uffe r 6S U BST IT U TIO N A SSA Y Use d t o d is tin g uis h f a c to r d efic ie n cie s ( d on e a ft e r o b ta in in g a n a b norm al A PTT O R P T) R EA G EN TS : N orm al p la sm a- A dso rb ed p la sm a- A ged s e ru m - Th ro m bin t im e- t e sts f o r F I d efic ie n cy A PTT-s c re en s i n trin sic a n d c o m mon p ath w ay P T- s c re en s e xtrin sic a n d c o m mon p ath w ay D efP T A PTT TT N orm al P la sm a Adso rb ed P la sm a Aged S e ru m IIIVV IIV IIIIXXX IXII

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