introduction to hydatid disease, diagnosis of hydatid disease,collection and transport of specimens,parasites,cysts

2. Learning Outcomes
Introduction to Hydatid Disease
Collection And Transport Of Specimens
Diagnosis Of Hydatid Disease

3. Introduction
 Human echinococcosis is a parasitic disease caused by larval stage of
tapeworms of the genus Echinococcus.(E.granulosus & E.multilocularis)
 E.granulosus causes cystic echinococcosis (hydatid cyst disease), which
is characterized by large slow-growing cysts, commonly in liver or lungs.
 Infective stage is the embryonated egg in feces & Diagnostic stage is
hydatid cyst in organs.
 Humans are infected through ingestion of parasite eggs in contaminated
food, water or soil, or through direct contact with animals.(zoonotic
disease)
 Remain asymptomatic for many years and symptoms may not present
until cyst or larval mass becomes large enough to produce local mass
effects.(0.3-1 cm per year)

4. Hydatid cyst
https://thebileflow.wordpress.com/
2012/10/31/pathology-hydatid-cyst

5. Collection of specimens
Blood(serum)
Hydatid cystic fliud
Urine

6. Preparation of serum
 serum
 Approxiatemetly 5 ml of venous blood collected from patient
whom had perfomed surgery transfered into red topped tubes.
 blood is collected under strict aseptic precautions and it is allow to
clot for 15-30 minutes at room temperature.
 Clot is removed by centrifuging at 1000-2000 x g for 10 minutes in
a refrigerated centrifudge to get the resulting supernatant
designated serum.
 Then, serum is immediately transfer into clean polypropylene
tubes/containers using pasteur pippetes.
 serum is alliquot,stored and transported at -20 or lower till use.

7. Handling and Transport Of Serum
Specimens
 Each container should be labeled with a code and date of collection.
 containers kept in non leakable zip lock bag and this bag should be placed
in another bag containing adequate packing materials(cotton) to absorb
liquid if leakage occurs.
 This package is placed in a thermocol box or small size cooler having ice
packs.( maintain proper cold chain system during transit)
 The box should be clearly labeled as “BIOHAZARD MATERIAL”.
 Routine laboratory request forms accompany each sample and specimens
should be sent to laboratory as soons as possible.

8. Materials and Equipment For
Transportation
 Transportation box
 Ice box
 Sample delivery register
 Disposable gloves
 Cotton/tissue rolls
 Zip lock bags
http://www.uvm.edu/safety/
lab

9. Method of collection
 Hydatid cystic fliud
 Percutaneous aspiration or Fine needle aspiration perfomed under
aseptic condition with continuos sonographic guidance.
 Skin above the lesions swabbed with antiseptic solution and the
patient is given a local anesthetic at the needle site.
 By using a transhepatic approach, the cyst is puncture by a sterile 18-
gauge needle and cyst contents aspirated rapidly into syringe.
 At the same time,warm gel will be applied on the target area and the
ultrasound transducer will precisely guide the needle into the cyst.
 ultrasound provides real-time visualization of the needle and cyst
during the procedure.
 After the needle is removed, a band-aid is applied to the needle site
and the fluid is transferred into cytolyt vials which will be transport to
the laboratory for analysis.

10. Diagnosis of
Cystic Echinococcus
 Diagnosis of cystic echinococcus is based on history of
epidemiologic risk factors,clinical evidence,radiology evidence and
serolological test.
 Radiodiagnosis ( ultrasound,CT scan,x-ray,MRI)
 Serological tests( ELISA,IHA,complement fixation,CO-A)
 Casoni's skin test(previously used)

11. Radiodiagnosis
(primary test)
 Ultrasonography
 it is a diagnostic imaging technique based on the application of
ultrasound. It is used to see internal body structures such as tendons,
muscles, joints, vessels and internal organs.
 cost-effective modality for examination of cystic lesions of liver.
 role of ultrasound in hydatid disease include first line diagnosis of
cysts,guided for precutaneous drainage and aspiration
prodecure(diagnosis & treatment)
 The 2001 World Health Organization (WHO) classification of hepatic
hydatid cysts is used to assess the stage of hepatic hydatid cyst based on
ultrasound and is useful for deciding the appropriate management,
depending on the stage of the cyst.

12. Ultrasound examination
(hydatid cyst in liver)
http://www.moleopedia.com/hy
datid-liver-disease

13. Classification of hydatid cyst
https://openi.nlm.nih.gov

14.  Computed Tomography(CT)
 A CT scan makes use of computer-
processed combinations of many X-ray
measurements taken from different
angles to produce cross-sectional
(tomographic) images of specific areas
of a scanned object inside body.
 plays a crucial role during the peri-
operative period for the detection of
complications such as cyst
rupture,underlying infection,billiary
and vascular movement.
 CT is an alternative if MRI is
unavailable.
 MRI allows anatomic and morphologic
characterization of lesions.( indicated
in CE if the cyst is in subdiaphragmatic
location,complicated by absecess)

15. Serological Test
(confirmatory)
 infection with echinococcus
induces an antibody response
most commonly IgG(IgG1,
IgG).
 A number of detection assay
are used to detect the
antibodies against hydatid
antigens.
 Serologic test such as Enyme-
linked immunoassay(ELISA) is
widely used because it has
sensitive methods for
http://www.microbiologynotes.com/elis
a-principle

16. Principle of ELISA In Cystic granulosus
 Echinococcus antigen(AgB) extracted
from hydatid cystic fliud of animal cyst
is bound on the surface of microtiter
strips.
 diluted sample serum from suspected
individuals are pipetted into the wells of
microtiter stripe.
 IgG antibodies in the serum will bind to
immobilized echinococcus antigen.
 after one hour incubation at room
temperature, the plate is rinsed with
diluted wash solution(to remove
unbound materials).
 then the anti-human IgG peroxidase
conjugate is added and incubate for 20
min.
 substrate (TMB) solution is added
inducing the formation of a blue dye in
wells. The colour formation of colour is
terminated by adding stop
solution.(blue to yellow)
 the resulting dye is measured and the
concentration of IgG antibodies directly
proptional to intensity of colour.

17. Principle Of Casoni's Skin Test
 casoni test is an immediate
hypersensitivity skin test
previously used in the diagnosis
of hydatid disease.
 Intradermal injection of 0.20ml of
hydatid fluid collected from
animal/human cyst which is
sterilized by seitz filtration.
 equal volume of saline(control)
injected on the other forearm and
observation made for next 30 min
and after 1 to 2 days.
 A wheal response occuring at
injection site considered positive
result.(immediate
hypersensitivity)
 As a precaution anaphylactic tray
must be kept ready before
carrying out the test.(Type 1
hypersensitivity reaction)
https://www.stepshealth.us

18. ACKNOWLEDGEMENT
I would like to grab this opportunity
to thank all the lecturers and my facilitator, Professor Dr.
Subhada Prasad Pani for their guidance and
support. Throughout the preparation and
presentation of this topic, i have gained and
broadened my knowledge. Thank you to all for
lending your ears.

19. References
 Imad S Dandan,Assaad M Soweid, MD, FACG, FASGE.Hydatid Cysts
Treatment&Management,Medscape.(Sep12,2016)http://emedicine.medscape.com/
article/178648-overview
 Clinicaloverviewchinococcosis,CopyrightElsevierBV.(March13,2017)https://www.cli
nicalkey.com//content/clinical_overview
 Echinococcosis Fact sheet,world Health Organization.(March
2017)http://www.who.int/mediacentre/factsheets
 Chaya, D., & Parija, S. C. (2014).diagnosis of cystic echinococcosis using
serum,urine,andcystfluidsamples.TropicalParasitology.https://www.ncbi.nlm.nih.gov
/pmc/articles