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Diagnosis of Leishmanisis ((with special emphases on DAT

: Presented by Tahani Alyas Epidemiology

TMRI

Leishmanisis* Most important vector-borne diseases * Protozoan parasites of genous leishmania* Transmited via a bite of Sand flies* Zoonotic Nature* Various Manifistations

Geographic Distrbution * Endemic in four Countenants* Occures in Africa,Asia,Middle East,Latin Amirica,Meditereian region and parts of Europe.* 350 milion at Risk* 1,5 milion cases Cutanious leishmanisis * 500.000 cases of Visceral leishmanisi

VL Cl Iran Bangladesh Brazil India Sudan Afghan India Syria Saudi Arabia Brazeil Beru

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AmazonIndiaNepalAfghan

Classification of Leishmania parasites

Kingdom : ProtistaPhylum : SarcomastigophoraClass : Zoomastigophora Order : KinetoplastidaFamily : Trypanosomatidae Genous : LeishmaniaSubg. : Leishmania ( Donovani, Tropica, infantum,etc)

Promastigotes of Leishmania

Amastigote of Leishmania

Leishmania Parasites and DiseasesDiseaseSPECIESCutaneous leishmaniasisLeishmania tropica*Leishmania major*Leishmania aethiopicaLeishmania mexicanaMucocutaneous leishmaniasisLeishmania braziliensisVisceral leishmaniasisLeishmania donovani*Leishmania infantum*Leishmania chagasi

The vectors ** Sand flies * Dipteran insects genus After sun set) * Nocturnal ( *70 out of 700 transmit leis- mania * Most known: P.aregntipes P. Orintalis P.papatasi

Sand fliesPhelobotomusLutzomiyia

reservoirIn rodentsIn carnivoresAnthroponotic (PKDL) Incriminated:More studies are needed)) Domestic animals

Reservior hosts

The life cycle of Leishmania

Clinical FormsCutanious Leishmanisis

* Orintal sore

* Skin ulcer

* Leishmania major , Leishmania tropica

* Severe form (Difuse cutanious leishmanisis) Leishmani athiopica, Leishmania amazonensis.

Clinical FormsMucocutanious Leishmanisis*known as espondia in new world*caused by:leishmania braziliansis (new world)L. athiopica in EthiopiaL. major & L. donovani in Sudan * Severe disfiguring of mucosal and carilignous tisuess of nose mutliation of face great suffering of life.

Visceral Leishmanisis

30Visceral Leishmaniasis or VL - is part of a family of diseases caused by the Leishmania protozoa. It is the most severe form of Leishmaniasis, with less severe forms affecting the cutaenous and mucotaneous tissue. Each form is caused by a different group of species of Leishmania, and there are over 20 species known to infect humans. VL is the second-largest parasitic killer in the world (after malaria), responsible for an estimated 100,000 deaths each year. Leishmaniasis is transmitted by a sand fly vector, and is endemic mainly in tropical regions, particularly the Indian subcontinent.

http://en.wikipedia.org/wiki/Visceral_leishmaniasis#cite_note-Dowell1997-38Visceral leishmaniasis: biology of the parasite Original Research ArticleJournal of Infection, Volume 39, Issue 2, September 1999, Pages 101-111Marcel Hommel17

Visceral Leishmanisis * Known as Kala-azar ( most severe form- fatal if left untreated)Caused by < L.donovani in Indian& East Africa < L.infantum in Mediterrenaean region < L.chagasi in New WorldReached internal organ ( liver,spleen and bone marrow)* 90% of cases in Bangladesh,Brazil, India,Nepal and Sudan.

PresentationFeverSplenomegaly, hepatomegaly, hepatosplenomegalyWeight lossAnaemiaCoughDiarrhoea

Diagnosis * Acuurate diagnosis is crucial ( effective treatment). * Preleminary diagnosis rely on clinical sighns * confitmative by serological or parasitological test

Parasitological It can be via < demonstration of parasite in relevant tissues(liver,spleen,lymphnodes and bone-marrow)

Common in VL)) < prepheral blood buffy

Parasitological (methods)Microscopic examination ( reliable-low sensivity).In vitro culture (promastigote in artificial media ( NNN,Tobies media). Senstive.Inculation of parasites in laboratory animals ( hamaster, mice, guinia pig).For routein). ( time cosuming, not suitable

ParasitologicalLeishmanin Test:Skin test(Montenegro)

Delyed type hypersensitivity Positive during Cl Negative in V

Molecular diagnosisMain detection based on : < PCR

< Hyperdization ( using brobe).

Serological Diagnosis

* Antigins Detection

* Antibody Detction

Ags Detection* LATEX Aglutination:< KATEX < Leishmani antigns in Urine< highly sensetive< Ags decrease after treatment

Ags Immunoblotting:

Recognize 14-110 kD in sera of VL

Antibody detection IFAT < detecting of circulating antibodies (IgG) ( human of CL&VL)< high ( senesitivty & specieficity)

Abs ELISA

< Detecting of Abs in VL:CL < Using whole ptromastigotes Ags< highly sensitive ( cross-reaction (Trypans, Tuber and Toxoplasma))< Highly specific ( using spesific L. Ags).

Abs detection K39