Aoun-Microsporidiosis 3rd EPU

7/31/2019 Aoun-Microsporidiosis 3rd EPU

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INTESTINAL MICROSPORIDIOSIS IN TUNISIA : G ROUPS AT RISK , P REVALENCE S & S PECIES IDENTIFICATION

CHABCHOUB N, ABDELMALEK R, A ISSA S, K ANOUN F, T IOUIRI H,BOURATBINE A & AOUN K


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Institut Pasteur de Tunis

Intestinal Microsporidiosis (Desportes, 1985)Emerging opportunistic infection (HIV infected +++)

Microsporidia pathogens: Protozoa / Fungi

1st descriptions in Tunisia in 1997: HIV patients, Congenitalimmunodefisciency & Immunocompetent child

Objective of the study:

Estimation of the prevalence of intestinal microsporidiaand identification of the species involved in Tunisia


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177 Immunocompromised patients:123 VIH infected & 54 Haematological malignancies (HIV -)

142 Healthy controls (children)

One Stool Sample

Studied Subjects


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279 pb

250 pb

250 pb E. bieneusi , 270 pb Enc. intestinalis , 268 pb Enc. cuniculi , 279 pb Enc. hellem

Universal PCR V1/PMP2 (Fedroko et al , 1995)Gene of the small sub unit rRNA

Sequencing


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E. bieneusi : V1/EB450(Zhu et al, 1993)

Enc. intestinalis : V1/IS500(Coyle et al, 1993)

353 pb375 pb

Species Specific PCRs


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Results


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Trichrome PCR pVIH (+)

(123)8 (6.5%) 18 (14.6%)

Prevalence in HIV patients

0,02

Prvalence in patients with Haematologic malignancies

Trichrome PCR p

VIH (-)(54)

0 5 (9.3%) 0,028

All Healthy Controls revealed negative


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Suggestive Size of amplified fragment (V1/PMP2): 22/23 (95.7%)

Sequencing: E. bieneusi L16868, E. intestinalis SIU09929, E. hellem L19070, E. cuniculi L17072 ( BLAST http:www.ncbi.nlm.nih.gov ) 19/23 (82.6%)

Specific PCR: E. bieneusi , Enc. Intestinalis 11/19 (57.9%)

Correlation in all cases

Species identification: 23 positive samples


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Species Proportion

E. bieneusi (n= 8) 42.1%

Enc. intestinalis (n = 10) 52.6%

Enc. hellem (n = 1) 5.3%


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PatientPCR (V1/PMP2) Sequencing

PCRsV1/EB450 V1/IS500

E. bieneusi Enc. intestinalis

R42 250 pb E. bieneusi -






R119 250 pb E. bieneusi - -

Sce1 250 pb E. bieneusi -

I14 270 pb Enc. intestinalis -

R60 270 pb Enc. intestinalis -

R92 270 pb Enc. intestinalis -

I44 270 pb Enc. intestinalis -

R68 270 pb Enc. intestinalis - -





I11 279 pb Enc. hellem - -

R15 270 pb 250 pb

Enc. intestinalis ND

--

+

-

R29 270 pb ND - -

R30 270 pb ND - -

I22 270 pb ND - -

A8 240 pb ND ND - -


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Stool aspect of HIV infected

Stool Patients Total

Diarrheal 7 (18.4%) 31 38

Molded 11 (12.9%) 74 85

Total 18 105 123

No statistical association

The infection was significantly associated with low T Lymphocyte CD4cell counts ( p=0,008 )

Clinical & Biological parameters


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Patient Age Stools Disease

I11 51 Diarrheal Myeloma


I22 ND Diarrheal Myeloma


A8 ND Molded BM aplasia

Haematological malignancies cases


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Genotyping of E. bieneusi ( 6 isolates): PCR-RFLP & Sequencing


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C ONCLUSION

High prevalence of Microsporidia infection inimmunocompromised patients in Tunisia

PCR much more sensitive than Microscopy

First microsporidia species identification in Tunisia:E. bieneusi & Enc. intestinalis; but also Enc. hellem

Co-transmission of Genotype B (Anthroponotic) et D(Zoonotic) of E. bieneusi


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JFMPMRabat 23-26 Oct 2013

Aoun-Microsporidiosis 3rd EPU

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