sepsissepsis
septicemia , anaerobic septicemiasepticemia , anaerobic septicemia
 
Timing of Blood CulturesTiming of Blood Cultures  ptimal time: !ust before anticipated onsetptimal time: !ust before anticipated onset
of the chill or fever,of the chill or fever,
Typically collected as soon as possible afterTypically collected as soon as possible after
thethe
  onset of fever or chills or "henever seriousonset of fever or chills or "henever serious
  infection is suspectedinfection is suspected
#ollection, "henever possible, prior to the#ollection, "henever possible, prior to the
administration of antimicrobial agents$administration of antimicrobial agents$
 Simultaneous collection of t"o to three setsSimultaneous collection of t"o to three sets
 
CulturesCultures && 'enipuncture is the method most 'enipuncture is the method most
commonly used(commonly used(
&&)henever possible, blood for culture)henever possible, blood for culture
should not be dra"n through anshould not be dra"n through an
ind"elling intravenous or intra*arterialind"elling intravenous or intra*arterial
catheter(catheter(
contamination+contamination+
Skin preparationSkin preparation
Poor skin preparation prior to dra"ingPoor skin preparation prior to dra"ing
bloodblood
cultures is the most common cause of culturecultures is the most common cause of culture
contamination$(contamination$(
&&alse*positive blood cultures may bealse*positive blood cultures may be
associatedassociated
"ith increased length of hospital stay and"ith increased length of hospital stay and
increased pharmacy and laboratoryincreased pharmacy and laboratory
charges$(charges$(
 
-./ alcohol for 0. sec from centre to periphery-./ alcohol for 0. sec from centre to periphery
1/ iodine tincture for 2. sec or 3./ povidone1/ iodine tincture for 2. sec or 3./ povidone
for 31. secfor 31. sec
-./ alcohol for 0. sec from centre to periphery-./ alcohol for 0. sec from centre to periphery
 4llo" to dry completely 4llo" to dry completely
Sterile glovesSterile gloves
Repalpitation of vein only "ith disinfected 5ngerRepalpitation of vein only "ith disinfected 5nger
#hange needle if unsuccessful#hange needle if unsuccessful
#leanse rubber stopper of the blood culture#leanse rubber stopper of the blood culture
 
 olume of Blood for olume of Blood for
CultureCulture The volume of blood dra"n per culture isThe volume of blood dra"n per culture is
the single most important variable inthe single most important variable in
recovering microorganisms from the bloodrecovering microorganisms from the blood
of bacteremic or fungemic patients($of bacteremic or fungemic patients($
&&6aboratories should routinely monitor the6aboratories should routinely monitor the
 volume of blood cultured as a quality volume of blood cultured as a quality
assurance activity7(assurance activity7(
8ach ml of blood,, up to 3. ml, can increase8ach ml of blood,, up to 3. ml, can increase
the sensitivity of the blood culture by 0 *the sensitivity of the blood culture by 0 *
9/$$9/$$
Optimal olume of BloodOptimal olume of Blood
for Culturefor Culture  4dults: 4dults: !" ml per set #t$o!" ml per set #t$o
bottles%bottles%
  * Infants and children* Infants and children !'( ml!'( ml
per bottleper bottle
setset
)umber of Blood)umber of Blood
Cultures SetsCultures Sets &&)ith adequate volume of blood, 1 *)ith adequate volume of blood, 1 *
0 blood culture sets are su%icient to0 blood culture sets are su%icient to
detect nearly all episodes ofdetect nearly all episodes of
bacteremia and fungemia(bacteremia and fungemia(
 
3; days for anaerobic3; days for anaerobic
; "eeks at least for brucella; "eeks at least for brucella
The duration is shortened "ith theThe duration is shortened "ith the
use of automated techniques$use of automated techniques$
 
Transport from Bed Side toTransport from Bed Side to
t*e Labt*e Lab Prior to transport, vials should be properlyPrior to transport, vials should be properly
identi5edidenti5ed
Transport time should be as fast as possibleTransport time should be as fast as possible
Transport temperatures should not be e<tremeTransport temperatures should not be e<treme
  preferably at RTpreferably at RT
  never fro=en nor refrigeratednever fro=en nor refrigerated
  not higher than normal body temperaturenot higher than normal body temperature
 'ial leakage should be considered all the time 'ial leakage should be considered all the time
  use adequate transport containersuse adequate transport containers
  "ear gloves"ear gloves
  4>T?4T8@ 4>T?4T8@
  ?4>46?4>46
Blood Culture +ediaBlood Culture +edia
o one medium or system is capable ofo one medium or system is capable of
detecting all microorganisms$detecting all microorganisms$
 4utomated systems o%ers special media 4utomated systems o%ers special media
for:for:
 ,utomated met*od ,utomated met*od
 'arious types of bottles available to isolate a range of organisms i$e$ aerobic, anaerobic,
 Mycobacterium
The system detects #arbon @io<ide production C this indicates presence of respiring organisms
 
co or metr c sensor anco or metr c sensor an
re-ected lig*t in Bactre-ected lig*t in Bact
 
in order to absorbin order to absorb
antibioticsantibiotics Resins are non*ionic and cationicResins are non*ionic and cationic
e<changers that neutrali=e many di%erente<changers that neutrali=e many di%erent
antibiotics "hich might be present due toantibiotics "hich might be present due to
the pretreatment of the patientthe pretreatment of the patient
help to lyse blood cells so that intra*help to lyse blood cells so that intra*
cellular organisms are set freecellular organisms are set free
provide the organisms "ith gro"th*provide the organisms "ith gro"th*
centres tocentres to
 
Biomerieu6 : 7emoline 4erformance DUO #7E+OLI)E DU
 
Dip*asic bottleDip*asic bottle
#ombination of:#ombination of:  4n agar slope covering one side of the bottle 4n agar slope covering one side of the bottle ;.ml of broth;.ml of broth
Both agar and broth contain gro"th factors, peptones, yeastBoth agar and broth contain gro"th factors, peptones, yeast e<tract, hemin, 4@ and vitaminse<tract, hemin, 4@ and vitamins
Broth contains anticoagulant ESPS: Sodium PolyanetholBroth contains anticoagulant ESPS: Sodium Polyanethol SulfonateFSulfonateF
 4tmosphere inside bottle is enriched "ith #arbon @io<ide 4tmosphere inside bottle is enriched "ith #arbon @io<ide and maintained at lo" pressure to create partial vacuumand maintained at lo" pressure to create partial vacuum
 4llo"s gro"th of main aerobic micro*organisms that 4llo"s gro"th of main aerobic micro*organisms that commonly cause sepsiscommonly cause sepsis
 
Dip*asic bottleDip*asic bottle
>sing>sing aseptic techniqueaseptic technique, the broth is inoculated, the broth is inoculated "ith 3.*31ml of blood and mi<ed"ith 3.*31ml of blood and mi<ed
The bottle is tipped to inoculate the agar slopeThe bottle is tipped to inoculate the agar slope Edo not tip blood into top of bottleFEdo not tip blood into top of bottleF
Bottle is incubated EuprightF for - days in totalBottle is incubated EuprightF for - days in total E; "eeks ifE; "eeks if Brucellosis Brucellosis is suspectedFis suspectedF
Bottle e<amined 1*0 times daily for colonialBottle e<amined 1*0 times daily for colonial gro"th on agar slope, broth turbidity,gro"th on agar slope, broth turbidity, haemolysis or a deposithaemolysis or a deposit
 
 ,naerobic bottle ,naerobic bottle
G.ml of broth enriched "ith gro"th factors,G.ml of broth enriched "ith gro"th factors,
peptones, reducing agents and addedpeptones, reducing agents and added
anticoagulant ESPS: Sodium Polyanethol SulfonateFanticoagulant ESPS: Sodium Polyanethol SulfonateF
 4llo"s gro"th of anaerobic micro*organisms that 4llo"s gro"th of anaerobic micro*organisms that
commonly cause sepsiscommonly cause sepsis
 4tmosphere inside the bottle is enriched "ith 4tmosphere inside the bottle is enriched "ith
#arbon @io<ide and#arbon @io<ide and HydrogenHydrogen and is maintained atand is maintained at
lo" pressure to create partial vacuumlo" pressure to create partial vacuum
Rubber stopper and red scre" cap present so easyRubber stopper and red scre" cap present so easy
to directly inoculate bottle using a needle and opento directly inoculate bottle using a needle and open
positive bottlespositive bottles
>sing>sing aseptic techniqueaseptic technique, the broth, the broth
is inoculated "ith 9ml of bloodis inoculated "ith 9ml of blood
and mi<edand mi<ed
@o not tip broth into lid of bottle@o not tip broth into lid of bottle
Bottle is incubated for 3; days inBottle is incubated for 3; days in
totaltotal
Bottle e<amined 1*0 times daily forBottle e<amined 1*0 times daily for
turbidity, haemolysis or a depositturbidity, haemolysis or a deposit
 
Blood culture bottles are asepticallyBlood culture bottles are aseptically
inoculated "ith blood samples and arrive ininoculated "ith blood samples and arrive in
laboratorylaboratory
@iphasic bottle is tipped to inoculate agar@iphasic bottle is tipped to inoculate agar
slopeslope
Incubate both bottles upright at 0-J#Incubate both bottles upright at 0-J#
Incubate aerobic bottles for - days in totalIncubate aerobic bottles for - days in total
E; "eeks ifE; "eeks if Brucellosis Brucellosis is suspectedFis suspectedF
Incubate anaerobic bottles for 3; days inIncubate anaerobic bottles for 3; days in
totaltotal
 
phasic bottle 8<amine agar slope for colonies8<amine agar slope for colonies 8<amine broth for turbidity, haemolysis or a deposit8<amine broth for turbidity, haemolysis or a deposit Subculture on Blood agar and ?ac#onkey agarSubculture on Blood agar and ?ac#onkey agar
Eeven if there is no gro"thF and incubate at 0-J# inEeven if there is no gro"thF and incubate at 0-J# in aerobic atmosphere for ;Ghours in totalaerobic atmosphere for ;Ghours in total
Subculture on #hocolate agar and incubate at 0-J#Subculture on #hocolate agar and incubate at 0-J# in #arbon dio<ide atmosphere for ;G hours in totalin #arbon dio<ide atmosphere for ;G hours in total
Perform Kram stainPerform Kram stain Perform direct antibiotic antifungal sensitivityPerform direct antibiotic antifungal sensitivity
testingtesting
Da3 !Da3 !
 4naerobic bottle: 4naerobic bottle: 8<amine broth for turbidity, haemolysis or a deposit8<amine broth for turbidity, haemolysis or a deposit Subculture to Blood agar and ?ac#onkey agar andSubculture to Blood agar and ?ac#onkey agar and
incubate in aerobic atmosphere at 0-J# for ;Gincubate in aerobic atmosphere at 0-J# for ;G hourshours
Subculture to #hocolate agar and incubate at 0-J#Subculture to #hocolate agar and incubate at 0-J# in #arbon @io<ide atmosphere for ;G hoursin #arbon @io<ide atmosphere for ;G hours
Subculture to Blood agar and incubate at 0-J# inSubculture to Blood agar and incubate at 0-J# in anaerobic atmosphere for at least ;G hoursanaerobic atmosphere for at least ;G hours
Perform Kram stain Toluidine bluePerform Kram stain Toluidine blue Perform direct antibiotic antifungal sensitivityPerform direct antibiotic antifungal sensitivity
testingtesting
pathogenspathogens
Perform appropriate identi5cation testsPerform appropriate identi5cation tests
e$g$ catalase, coagulase, o<idase, grame$g$ catalase, coagulase, o<idase, gram
etcetc
If direct sensitivities are poor, performIf direct sensitivities are poor, perform
antibiotic antifungal sensitivity testingantibiotic antifungal sensitivity testing
on organismEsF gro"n on subcultureson organismEsF gro"n on subcultures
 
4ossible contaminants4ossible contaminants Blood does T have a normal microbial LoraBlood does T have a normal microbial Lora  4septic technique 4septic technique  ?>ST be used "hen taking?>ST be used "hen taking
blood sample, adding it to the bottles and "henblood sample, adding it to the bottles and "hen processing positive bottles in the laboratoryprocessing positive bottles in the laboratory
#ommon environmental organisms include:#ommon environmental organisms include:  Bacillus Bacillus speciesspecies  Acinetobacter Acinetobacter speciesspecies
#ommon skin microbial Lora include:#ommon skin microbial Lora include: #oagulase*negative#oagulase*negative staphylococcistaphylococci  'iridans 'iridans streptococcistreptococci  Micrococci Micrococci CorynebacteriumCorynebacterium speciesspecies
    contamination of bloodcontamination of blood
culture bottlesculture bottles Sterilise patients skin "ith alcohol "ipe or iodine solution beforeSterilise patients skin "ith alcohol "ipe or iodine solution before taking blood sampletaking blood sample
>se a sterile needle to take sample>se a sterile needle to take sample The needle should not touch any other surface before sample is takenThe needle should not touch any other surface before sample is taken The top of the bottles should be sterilised "ith alcohol "ipe or iodineThe top of the bottles should be sterilised "ith alcohol "ipe or iodine
solution before sample is addedsolution before sample is added The needle should not touch any other surface before adding sampleThe needle should not touch any other surface before adding sample
to bottlesto bottles The top of the bottle should be sterilised "ith alcohol "ipe or iodineThe top of the bottle should be sterilised "ith alcohol "ipe or iodine
solution after adding samplesolution after adding sample )hen tipping the bottle to inoculate the agar slope do not tip broth)hen tipping the bottle to inoculate the agar slope do not tip broth