In this slide i have discussed all the conventional tools to diagnose parasitic infections and also included all the new diagnostic advances have came to deep tech world.

1. Presented by-
ARPITA CHANDRA

2. ACKNOWLEDGEMENT
First and foremost, I would like to thank for,
Dr.Subhash Chandra Parija, Vice Chancellor, Shri Balaji
Vidyapeeth from Puducherry given such a knowlegable
Lecture in online series based on “Basic Principles of
Laboratory Diagnosis of Parasitic Infection”.
My special thanks gives to Dr. R. Sujatha, Professor and
Head of the Department, for giving me such a
opportunity to present it.
I m also thankful to all my Teachers of the Microbiology
department.

3. Introduction
In 2023, the World Health
Organization (WHO) reported
an estimated 137,000 deaths,
caused by foodborne parasitic
diseases.
Vector-borne diseases caused
by parasites every year, with
400,000 being caused by
malaria alone.

4. Amoebiasis is an intestinal parasitosis
caused by the protozoan Entamoeba
histolytica that represents the third
leading cause of mortality due to
parasitosis.

6. Types of Examination Method Names
Gross Gross examination
Microscopic
Direct wet smear
Concentration technique
Staining technique
Scotch tape technique
Skin scraping technique
Culture
Harada–Mori technique
Baermann technique
Charcoal culture technique
Agar plate culture technique
Immunological
Enzyme-linked immunosorbent assay
(ELISA)
Immunoblot assay
Molecular biology
Polymerase chain reaction (PCR)
Loop-mediated isothermal amplification
(LAMP)
xMAP assay (Luminex)
Restriction fragment length
polymorphism (RFLP)
Next-generation sequencing (NGS)
Mass spectrometry (MS)
Biosensors

7. Gross / Macroscopic Examination
 Consistency
 Color
 Odor
 Presence of blood
 Presence of mucus
 Presence of adult worms or segments
of tapeworm

8. Microscopic
examination

9. Concentration Technique
Floatation technique
- Saturated sodium chloride technique
- Zinc sulfate technique
Sedimentation technique
- Simple sedimentation technique
- Formal ether technique
- Formalin acetone technique

11. 3.Staining techniques
Iodine mount
For Nuclear details of cysts, helminthic eggs and Larvae
Advantage- Better visualization
- Species identification
Disadvantages - Iodine immobilizes and kills parasite
- Lowe sensitivity (25-60%)
- can not appreciate for
Motility of the protozoan trophozoites &
Helminthic larvae

12. Types of Iodine stains
 Lugol’s Iodine
 D’Antoni iodine
 Dobeil’s iodine
2. Permanent stained smear
(Accurate diagnosis, stain for internal structure)
1. Iron hematoxylin stain
2. Trichrome stain
3. Modified acid fast stain (Cryptosporidium,
Cyclospora & isospora belli)

13. 4. Scotch tape technique
(Use For diagnosis of pinworm) – Enterobius
vermicularis
5. Skin scrapping technique
(Use for diagnosis of Ectoparasites)- e.g.
Sacroptes, Otodectes.

14. CULTURE
Culture techniques Uses
Harada Mori
technique
Strongyloides
larvae
Baermann
technique
Identification of
3rd stage Larvae
( based on active
movement of
larvae)
Charcoal
culture tech
Infective stage of
Strongyloides
larvae
Agar plate
culture tech
S. Stercoralis
and Hookworm

15. Fig - Timeline of technologies for medical parasite and arthropod
diagnoses. Microscopy has been applied for pathogen and insect
identifications for several centuries. To improve detection efficacy, a
range of techniques have emerged. They encompass artificial
intelligence, ELISA, PCR, geometric morphometrics, and DNA barcoding
technique

16. Immunological Examination
ELISA
IMMUNOBLOT ASSAY
Use for Detection of
Ag/Ab of Ameobiasis,
Chagas’ disease,
Toxoplasmosis,
Schistosomiasis and
Lymphatic filariasis
Useful for Diagnosis
of schistosomiasis,
strongylodiasis, and
hydatid disease
Rapid card test
e.g.- Malaria parasite

17. Advantages of Antigen detection tests
 Detect circulating parasite Ag/Ab in serum, urine,
genital specimens or feces
 Detect acute/recent infection
 Monitor response to treatment
 Useful when microscopy is negative.
LIMITATIONS
-high cross-reactivity,
-time consuming,
-labor intensive,
-unable to detect low-level antibodies
-costly reagents
Higher sensitivity
(80 to 94%)
Specificity
(94 to 100%)
than microscopy
and culture

18. Shift in parasite detection strategy landscape

19. Molecular Biology
1. DNA Probe
Radiolabelled neucleotide sequences
Complementary to a part of the parasite
(clinical samples)
Use for P. falciparam, W. bancrofti

20. 2. Polymerase Chain Reaction (PCR)
The Operational framework in molecular diagnostics

21. Most useful for targeting 16S- rRNA gene for
differential detection of all the three morphologically
similar forms of
E. histolytica, E. moshkovskii and E. dispar
simultaneously in stool samples

22. Entamoeba
 Pseudopod forming protozoan
parasites
 Possess nucleated cysts(larger-
8neuclei)
 7 different species (E. histolytica,
E. coli, E. hartmanni, E. polecki,
E. dispar, E. moshkovskii and E.
bangladeshi)-Human Intestine
 E. gingivalis – Oral cavity
commensal
 SOI-Contaminated Food and
water

23. Most widely used PCR marker for taxonomic
differentiation between E. histolytica, E. dispar
and E. moshkovskii.

24. Molecular diagnosis of amoebiasis in stool sample
Fig. Complete Process of Nested multiplex PCR for differential
detection of Entameoba
STOOL
SAMPLE

25. Some approachs in molecular diagnosis of amoebiasis


26. Molecular diagnosis of amoebiasis in stool
samples

27. Molecular diagnosis of ameobiasis in urine
samples

28.  Running cost (consumables & equipment)
 Requirement of laboratory setup
 Unavailabilty of detection kits for many parasites
 Trained technical manpower
These are acting as the
impediment for the bench-to-field
of the technology especially for
the lower and Middle Income countries

29. Move to Deep Technology Era of Parasitic
Diagnosis

30. What is DEEP TECHNOLOGY
- Those techniques whose working model is based on
high tech innovation in scientific advances
-Come up with innovative solutions
-Approaches to adressing big societal challenges

31. What Deep Tech will offer ?
 Maximum sensitivity
 Maximum specificity
 Portability
 Scalability
 User friendly
 Multiple parasitic detection
 Parasite stages detection
 focusing on publicly available microscopic image
datasets of protozoan parasites.

32. Diagnosis of parasitic infection
by mobile phone ?

33. IIT Kharagpur faculty develops smartphone app for
Malaria in 2015
Malaria screener
First Smartphone-based system
Automated malaria light microscopy
Screen thin and thick smears
Fig- System Setup for Malaria Screener.

34. Malaria Parasite detection from Blood Smear
WORKFLOW

36. Suspect Malaria – Use you
Smartphone !

37.  Dataset contains four species of Malaria
parasites: Falciparum, Malariae, Ovale, Vivax
 Six types of protozoan parasites
 (Plasmodium, Toxoplasma, Babesia, Leishmania,
Trypanosome, Trichomonad)
 Host cells (RBCs and Leukocyte)
TARGETS

38. Not only Parasite, the Deep Tech also enable
identification of stages
Sci
Rep 11,16919

39. Why only one Parasite
Lets detect all in sample
VETSCAN
IMAGYST
ParasiteVectors 13,346 (2020)

40.  Requires reference data sets
from database
 Still limited to some parasites
 Need for re-imagination
 Need to push science
boundaries
 Difficulties in scaling up
 Difficulties in accessing
funding

41. Conclusion
Microscopy with poor sensitivity and
Serological diagnosis have also several limitations……BUT
Molecular studies is useful tool for understanding pathogenesis, help
developing novel treatment strategies !
Deep Tech innovations offers disruptive solutions
Zero interpretation biases, Detect even single
parasite