NRL for Diagnosis of Parasitic Diseases

All parasitological laboratories and clinical departments in the country are provided with technical assistance. NRLDPD is also in help of the MH and Regional Health Inspectorates in cases of outbreakes and epidemics. Epidemiological work and annual analysis of the parasitic morbidity is conducted as well as evaluation of the activities of the parasitological network in the country. The laboratory staff is involved in postgraduate education.

The laboratory carries out routine and reference diagnosis by applying contemporary laboratory methods – morphological, immunological and biomolecular.

1. Intestinal helminthic diseases: Аscariasis, Trichuriasis, Enterobiasis, Тaeniasis, Hymenolepiasis, Strongyloidiasis, Ancylostomiasis, Diphyllobothriasis, Fascioliasis, Dicrocoeliasis, intestinal Schistosomiasis and other rare or tropical helminthic diseases. Material: stool
   • Маcroscopic examination and morphological differentiation of intestinal helminths
   • Microscopic examination for helminth eggs and larvae:
     • Wet mount preparations – with saline and iodine solutions
     • Concentration procedures (sedimentation, flotation, formalin - ethyl acetate sedimentation technique)
     • Detection and larvae cultivation - Baermann funnel technique, Harada-Mori filter paper technique
     • Transparent tape test: Enterobiasis and Taeniasis (T. saginata)
   • Immnodiagnostic methods in Fascioliasis (ELISA, IHA, IFA)
2. Intestinal protozoal diseases (Giardiasis, Amebiasis, Balantidiasis, Blastocystis infection, Cryptosporidiosis, Cystoisosporiasis, Cyclosporiasis, Мicrosporidiosis). Materials: stool, duodenal material (Giardiasis), biopsy material:
   • Мicroscopic examination for trophozoits, cysts and oocysts:
     • Wet mount preparations – with saline and iodine solutions
     • Concentration procedure – formalin - ethyl acetate sedimentation technique
     • Staining procedures – Giemsa, trichrome, modified Ziehl Neelsen staining, etc.
   • Cultivation: Pavlova’s medium (Аmebiasis, Blastocystis infection)
   • Immunodiagnostic methods in Amebiasis (ELISA)
   • Biomolecular methods: PCR for Giardiasis, Blastocystosis, Cryptosporidiosis
3. Blood and tissue parasitic diseases

   Мalaria and Babesiosis. Materials: peripheral (capillary) blood for thick and thin blood films, Rapid Diagnostic Tests and PCR

   • Microscopic examination: Staining procedures - Giemsa stain
   • Biomolecular methods: PCR for malaria species identification

   Visceral leishmaniasis. Materials: bone marrow and serum for immunodiagnostics, as well as biopsy materials from lymph node, liver, and spleen.

   • Microscopic examination:
     • Staining procedures - Giemsa stain
   • Cultivation techniques: cultivation in Novy-MacNeal-Nicolle (NNN) medium
   • Immunodiagnostic methods – ELISA, Western blot
   • Biomolecular methods – PCR for diagnostics and species identification

   Cutaneous and mucocutaneous leishmaniasis: Material: skin lesion biopsy specimens:

   • Microscopic examination:
     • Staining procedures – Giemsa stain
   • Cultivation technique: cultivation in Novy-MacNeal-Nicolle (NNN) medium

   Cutaneous amebiasis. Material: bipsy specimen from skin lesion:

   • Microscopic examination:
     • Staining procedures – trichrome
     • Examination of histological material
   • Cultivation technique: cultivation in Pavlova’s medium

   Toxoplasmosis. Materials: biopsy specimens - lymph node, spleen, liver, intraocular fluid, placenta, amniotic fluid, as well as cerebrospinal fluid and serum for immunodiagnosis:

   • Microscopic examination:
     • Staining procedures – Giemsa stain
     • Examination of histological materials
   • Immunodiagnostics: ELISA (IgG, IgM, IgA antibodies, ELISA – IgG avidity), Western blot
   • Intraperitoneal inoculation into mice
   • Detection of parasite DNA by PCR

   Trypanosomiases (American and African). Materials: venous blood, biopsy material, cerebrospinal fluid

   • Microscopic examination:
     • Staining procedures – Giemsa stain
     • Examination of histological materials

   Extraintestinal amebiasis. Material: serum, skin lesion biopsy specimens

   • Immunodiagnostics: ELISA, IFA

   Naegleria and Acanthamoeba infections. Materials: cerebrospinal fluid, biopsy specimens (brain tissue, skin, cornea) or of corneal scrapings

   • Microscopic examination:
     • Wet mount preparations
     • Staining procedures - trichrome, Heidenhain, Lawless stains
     • Examination of histological materials
   • Cultivation technique – non-nutrient agar (NNA) plates, PPG medium
   • Моlecular methods - PCR

   Pneumocystosis. Materials: induced sputum, bronchoalveolar lavage, biopsy specimen

   • Microscopic examination:
     • Staining procedures – Giemsa, toluidine blue

   Еchinococcosis. Materials: serum sample, pathological specimens for detection of present scolexes in the cyst and vitality of the parasitic cyst:

   • Microscopic examination:
   • Examination for presence of parasitic elements – hooks, scolexes and membranes
   • Methylene-blue staining for vitality determination
   • Immunodiagnostics: ELISA, Western blot

   Toxocariasis (Toxocarosis). Material: serum sample:

   • Immunodiagnostics: ELISA, Western blot

   Тrichinellosis (Trichinosis). Materials: serum sample, muscle biopsy specimen, suspicious meat products for sanitary examination:

   • Microscopic examination:
     • Examination for presence of trichinella larvae
   • Immunodiagnostics – IHA, ELISA, Western blot
   • Molecular analysis and Trichinella genotype determination – Multiplex and Nested PCR
   • Sanitary examination for presence of Trichinella larvae in meat products
     • Compressive trichinelloscopy (Squash preparation)
     • Digestion with artificial gastric juice

   Cysticercosis. Materials: serum sample, cerebrospinal fluid (CSF), and biopsy histological specimens

   • Microscopic examination:
     • Examination of hystological specimens for parasitic elements
   • Immunodiagnostics - ELISA

   Paragonimiasis. Materials: bronchoalveolar lavage, tracheobronchial secretion, induced sputum, biopsy specimen, stool

   • Microscopic examination:
     • Wet mount preparations
     • Concentration techniques

   Filariases. Materials: venous blood, biopsy specimen

   • Staining procedures – Giemsa stain
   • Concentration techniques: 2% formalin (Knott's technique)
   • Examination of subcutaneous nodes (Onchocerca volvulus and Mansonella streptocerca)
   • Rapid diagnostic testMicroscopic examination:

   Dirofilariases. Material: biopsy specimen

   • Мicroscopic examination:
   • Examination of histological specimens
   • Маcroscopic examination, after parasite extraction
4. Urogenital parasitic diseases

   Тrichomonasis. Materials: urine, semen, prostate exprimate, vaginal and cervical swab specimens, urethral swab specimen:

   • Microscopic examination:
     • Wet mount preparations
     • Staining procedures – Giemsa stain
   • Cultivation techniques: cultivation in TV4 medium
   • PCR

   Schistosomiasis (urogenital). Materials: 24 hours urine, urine collected in the interval between 11.00 and 15.00 o’clock – after physical exercise, bladder biopsy specimen:

   • Microscopic examination:
     • Wet mount preparations
     • Concentration techniques – sedimentation, formalin - ethyl acetate sedimentation technique
     • Rapid diagnostic test – with urine

Members of the laboratory participate in the implementation of the following scientific projects financed by the Scientific Research Fund: