Presentation is loading. Please wait.

Presentation is loading. Please wait.

Chagas disease in Spain: Experience from a reference laboratory

Similar presentations


Presentation on theme: "Chagas disease in Spain: Experience from a reference laboratory"— Presentation transcript:

1 Chagas disease in Spain: Experience from a reference laboratory
Second WHO consultation: Development of a WHO reference panel for the control of Chagas diagnostic tests Geneva, 27 – 28 January 2009 Chagas disease in Spain: Experience from a reference laboratory Teresa Gárate Servicio de Parasitología Centro Nacional de Microbiología

2 Experience from a Parasitology Reference Laboratory
General Role of the Parasitology Lab Parasitology Lab and Chagas’ disease in Spain

3 Parasitic diseases National Microbiology Centre AUTOCTHONOUS IMPORTED
Leishmaniasis Toxoplasmosis Cryptosporidiasis Giardiasis Amebiasis Fasciolosis Hydatidosis Anisakiasis Toxocariosis Trichinellosis IMPORTED Malaria Other Leishmaniasis Chagas disease Sleeping sickness Cysticercosis Schistosomiasis Filariasis/Oncho Intestinal parasites National Microbiology Centre

4 The Parasitology Lab works with Spanish hospitals and blood donor centers on Chagas disease diagnosis

5 Other Activities related with T. cruzi diagnosis
Chagas Test evaluation In house tests Commercial tests Control sample preparation Serum of characterized patients DNA from parasite strains Blood spiked with parasites Training microbiologists in Microhaemotocrit PCR Serological result interpretation

6 Implementation of T. cruzi Diagnostic Tools
in Parasitology Lab Commercial assays (rapid tests) PCR ELISA Microscopic examination Xenodiagnosis IFAT Culture Real time PCR

7 Evaluation of serological tests: In house ELISA and 3 serum panels

8 Evaluation of serological tests: In house ELISA and 3 serum panels
Diagnostic index Panel 1 Panel 2 Panel 3 % CI 95% Sensitivity 85 65 to 100 97 91 Specificity 98 96 PPV 94 NPV 88 73 99

9 Characteristics of serum samples of the 3 panels used in the evaluation of the In house ELISA
Panel 1: Sera from Bolivian individuals with positive or negative xenodiagnosis Panel 2: Sera from population with positive or negative T. cruzi IFAT. Healthy individuals from endemic and non-endemic areas Panel 3: Sera of anti-T. cruzi Specific Panel and healthy individuals from endemic and non-endemic areas Anti-T. cruzi Specific Sera Panel (QPanel, Sao Paulo-Brazil, was kindly provided by Dra. Elena Franco

10 Evaluation of serological tests:
Commercial ELISAs

11 Characteristics of serum samples used in the evaluation of commercial tests

12 Evaluation of Commercial ELISAs: Sensitivity

13 Evaluation of Commercial ELISAs: Specificity
a The specificity was calculated into account results of healthy control b The specificity was calculated into account results of healthy control and other pathologies.

14 Molecular diagnosis of T. cruzi
Control assay of limit detection Samples of population at-risk kDNA-PCR modified ( / HUF-REV) Britto et al 1993; Cruz et al 2002; Walsh et al 1991; Dorn et al 1997; Gomes et al 1998; Wincker et al 1994; Rubio et al 2002

15 Molecular diagnosis of T. cruzi Diagnostic sensitivity and specificity
PCR Positive Negative Total Acute patients 11 Chronic patients 471 295 766 Seronegative subjects 1227 482 1522  2004 Diagnostic index in chronic phase CI (95%) Sensitivity 61.5 58 65 Specificity 100 99.96 Positive predictive value 99.89 Negative predictive value 80.62 78.6 82.64

16 Chagas disease cases in Spain
AUTOCTHONOUS Blood transfusion Congenital transmission Organ transplant Laboratory accidents IMPORTED Immigration from endemic area Long stays in endemic area

17 Latin American population in Spain On December 2008: > 1.8 million

18 Reports of Transfusional Chagas cases in Spain
Year of detection Year of Donation N º of recipients Nº of infected Ref. 1992 ? 1? 1 Villalba et al., 1992 2005 2004 1 Fores et al., 2007; Flores-Chavez et al., 2008 2006 10 1 Abalo et al., 2007, CNM 2007 9 2 Perez et al., 2008; CNM 2008 2004 1 Ibarra et al., 2008, CNM National Haemovigilance System

19 Transfusional Chagas Cases
Madrid recipient Coruña recipient Málaga recipient 1 Málaga recipient 2 País Vasco recipient Origin Spanish Moroccan Age 27 55 33 57 4 Clinical status before transfusion Leukemia Hepatic transplant Bone marrow aplasia Choroid plexus papilloma Cardiac surgery Phase of Chagas disease and symptoms Acute FUO Multiorganic failure Chronic Asymptomatic Right bundle branch block Treatment Yes Follow up After treatment Fatal dead person S (+) PCR (-) S (-)

20 Characteristics of Infected donors:
País Vasco A Coruña Madrid Characteristics of Infected donors: Malaga A Coruña Madrid Malaga País Vasco Origin Bolivia Brazil Sex Male Female Age 62 58 53 ELISA 2.3 2.2 IFAT > 1/160 PCR P N/P Treatment No Yes Permanence in Spain 42 years 1 year 3 years

21 Congenital Chagas disease Newborns examined during 2002 - 2008
Positive PCR 2002 1 2003 3 2004 2 1* 2005 4 2006 12 2007 78 2008 91 5 Total 191 9 Transmission index: ~ 4,5% * Post mortem diagnosis In Spain ≈ women of childbearing age Bolivia: women Argentina: women

22 Laboratory Accidents and Chagas disease
Alvar J Laboratorio 76(456):

23 Chagas disease cases in Spain
AUTOCTHONOUS Blood transfusion Congenital transmission Organ transplant Laboratory accidents IMPORTED Immigration from endemic area Long stays in endemic area

24 C.C. de Sangre y Tejidos de Asturias
Chagas disease in blood donors at- risk Asturias Madrid C. T. Cruz Roja de Madrid Prevalence % 1,23 C.C. de Sangre y Tejidos de Asturias 0,13

25 Profile of immigrant population
Madrid Asturias 2006: ≈ immigrant 2008: ≈ immigrant 2006: ≈ immigrant 2008: ≈ immigrant

26 Prevalence in blood donors at-risk (2002-2006)
P N Prevalence % Nicaragua 1 16 6.25 Colombia 776 0.13 Venezuela 249 0.40 Ecuador 908 0.11 Peru 2 258 0.78 Bolivia 37 203 18.23 Brazil 221 0.90 End. Zone 3 207 1.45 Non End. Zone 307 0.33 ND 245 1.22 Spain 277 0.36 ? P = Seropositive N = Seronegative

27 Seroprevalence in population at-risk: 24%
≈ 1144 Chagasic patients

28 Distribution of cases in Spain

29 Origin of seropositive population

30 Conclusions Panel of Reference
- Positive samples by different serological tests, and - Positive samples from chagasic individuals with parasitological, epidemiological and clinical evaluation. Number of positive sera/panel - 3 serum samples for kit manufacturers - 100 serum samples for evaluation of tests - 3 serum samples for quality control. Chagas in Spain - Imported and autochonous types. - Bolivian group, mainly from Santa Cruz region, shows the highest seropositivity rates and represents the highest risk for T. cruzi infection transmission.

31 Thank you UNIDAD LEISHMANIA Y CHAGAS Carmen Cañavate Javier Nieto
Mercedes Rodriguez Israel Cruz Elena Bodas Marta Hernández Rubén González Emilia García Carmen Chicharro María Flores

32 ? Chagas disease in Spain AUTOCTHONOUS IMPORTED
Transfusional and congenital Chagas Immigration and long stays in endemic area Organ transplant and lab incident ? Months Years Acute Chronic phase

33 Level of anti-T. cruzi IgG in samples of patients with positive PCR
Acute case 1 (Madrid recipient) Acute case 2 (Málaga recipient)


Download ppt "Chagas disease in Spain: Experience from a reference laboratory"

Similar presentations


Ads by Google