Bartonella (2003-2007).

P. Ventosilla, Sandra Palma, Sandra Villar, Marcos Gonzales,

J. Chauca, H. Guerra,

C. Maguiña.

OBJECTIVE

To development immunological tests for diagnosis, research and

transfer to the community (2003-2007).

Perú

Ecuador

Colombia

A M A Z O N A S

C A J A M A R C A

A N C A S H

L I M A

AMAZONASPIURA

CAJ AMARCASAN MARTIN

HUANUCOANCASH

LIMA

AYACUCHO

AMAZONASPIURA

CAJ AMARCA

SAN MARTIN

LA LIBERTAD

ANCASH HUANUCO

LIMAJ UNIN

CUSCO

LORETO

AM

AZO

NA

S

PIURA

HUANUCO

CUSCO

1995 1999 2001

2005

BARTONELLOSIS REPORTED BY BARTONELLOSIS REPORTED BY DEPARTMENT IN PERUDEPARTMENT IN PERU

1997

MINSA/OGE- 20062006

3258 cases12 dead12 departments

Before DIGOS Support:

Birtles R, Fry N, Ventosilla P, Cáceres A, Sánchez E, Vizcarra H, Raoult D.J. Clin Microbiol 2002; 40:3606-12 Identification of Bartonella bacilliformis genotypes and their relevance to epidemiological

investigations of human bartonellosis.

Rolain JM, Novelli S, Ventosilla P, Maguina C, Guerra H, Raoult D. Immunofluorescence detection of B. bacilliformis flagella

in vitro and in vivo in human red blood cells as viewed by laser confocal microscopy. Ann N Y Acad Sci. 2003 Jun;990:581-4

Act. 6.5.1.1. To standardize western blot test for research and diagnosis of bartonellosis.

Act. 6.5.1.2. To standardize ELISA technique for research and diagnosis of bartonellosis; selection of the antigens from B. bacilliformis.

Act. 6.5.1.3. To Standardize DOT-Blot technique for research and diagnosis of bartonellosis, compare the sensibility and specificity with Western-blot, ELISA-Pilot study in a bartonellosis endemic area (2006-2007).

Act 6.5.1.4. Transfer of deliverables to the health system and Community (2007).

PA= Pooled acute phase seraPV= Pooled chronic phase sera PN= Pooled negative control sera

Results: Act 6.5.1.1. Western Blot:

The preparation of antigen by sonication revealed 26 protein bands (2.5-132 kDa); 3 antigenic bands (8 - 70 kDa) were recognized by acute phase sera and 6 bands by chronic phase sera (10 kDa – 70 kDa).

Results: Act 6.5.1.1. Western Blot using individual Protein Antigens (Bands):

M. W. Band

Sensitivity Specificity

65 kDa 76.3 %(29/38)

87.5 % (28/32)

12 kDa 42,1 % (31/36)

100 %(32/32)

13 kDa 26.3 %(10/38)

100 % (32/32)

M. W. Band

Sensitivity Specificity

65 kDa 86.1 % (31/36)

87.5 % (28/32)

12 kDa 88.8 % (32/36)

100 % (32/32)

13 kDa 41.6 % (15/36)

100 % (32/32)

Table 1. Sensitivity and specificity in the acute phase

Table 1. Sensitivity and specificity in the chronic phase

• In acute phase patients: the sensitivity was 82% and the specificity was 70%.

• In chronic phase patients: the sensitivity was 91% and the specificity 76%.

Results: Act 6.5.1.2. ELISA

• Sera: 1/50

• Conjugate:1/300

• Crude Antigen: 2mg

Results: Act 6.5.13.: Standardization of DOT-BLOT

Act 6.5.1.4. Transfer of deliverables to the health system and Community

(2007)

We will have transferred the

technology at health system and

community by the end of the 2007.

Products: In technology

• To have Western blot and Elisa with a good sensibility and specificity.

• We will have obtained a dot-blot by the year 2007.

• We have 940 sera, 110 plasma, 32 strains of B. bacilliformis from different part of the endemic area.

• We have a media to produce antigens and criopreserve.

Products: Human resources strengthening

Young Researcher in Bartonelosis: we were tutorof 4 international students (Holland, USA,Germanyand Canada).Thesis: • 3 students will have obtained an a Licentiate in Biology

from Universidad Nacional Federico Villarreal, by the year 2007.

• 1 student from UPCH will have obtained a PhD degree by the year 2007.

• 2 students will have obtained a degree of medical technology in UPCH, by the year 2007.

Products: Human resources strengthening

Training: We havetrained in clinicaland laboratorydiagnosis ofBartonelosis about115 persons from theHealth Minister andnon governmentalinstitutions.

Products• We were consulted for the Bartonelosis from national authorities as INS,

EO, MH that leading to health policy changes.

• We have participated in technical meeting for the validation of the norms and Procedures for the control of the Bartonella in Peru. Main directorate of Health, Direction of the Control Program of Transmissible Diseases, Ministry of Health. Lima.

• We received invitations for scientific societies as ASR and training activities, as well as I had been invite to be member of American Society for Rickettsiology.

• We received invitations for research grant proposals from Researcher of USA and Liverpool University.

• We are involvement in national and international scientific networks of Bartonelosis and Rickettsiology.

Products 4 Abstracts at International Congress were present:• Huarcaya E ; Maguiña C ; Best I ; Solorzano N ; Menacho J Ventosilla P.

Immunological pattern of patients with acute and chronic phase of Bartonella bacilliformis infection in a endemic area in Peru. 55th Annual Meeting the American Society of Tropical Medicine and Hygiene (55° nov.12-16 : Atlanta, Georgia. United States) In: The American Journal of Tropical Medicine and Hygiene: nov. 2006: 75(5) supl.(Abstract 1066)

• Ventosilla, Palmira; Maguiña, Ciro; Best Ivan; Villar Sandra; Palma Sandra; Henriquez César; Huacaya, Erick. Travel Award for presenting research findings at a the 20th meeting of the American Society for Rickettsiology And the 5th International Conference on Bartonella as Emerging Pathogens in Pacific Grove, Califronia, USA, September 2-7 2007.

• S. Palma, M. Villar, M. Gonzáles, C. Prause, C. Maguiña, P. Ventosilla. Two immunological test to diagnostic carrion´s disease (bartonellosis) in patients of endemics areas in Peru. ECCMID-ICC meeting to be held in Munich from 31 March to 3 April 2007. Manuscript in preparation

• R. M. Castillo, C. Terrones, D. Yabar, P. Ventosilla. Knowledge, Attitudes and Practices regarding Bartonellosis of the residents of Ollantaytambo district, in the Urubamba province, in the Sacred Valley of the Incas, Cusco-Peru, July 2004. In the 17th Latin American Symposium, 73rd AMCA Annual Meeting, Orlando Florida, April 1-7, 2007.

Publication

12 publications:• Compromiso Inmunológico en Pacientes con

Bartonelosis Humana. E. Huarcaya, I. Best, J. Merello, C. Maguiña, P. Ventosilla. Aceptado para la publicación por Fundación Instituto Hipólito Unánue, en Diagnóstico.

• Report of an unusual case of persistent bacteremia by bartonella bacilliformis in a splenectomized patient. Henriquez C, Hinojosa JC, Ventosilla P, Infante B, Merello J, Mallqui V, Verastegui M, Maguina C.Am J Trop Med Hyg. 2004 Jul;71(1):53-55.

• Chapter Bartonella, In Dermatology, C. Maguiña, H. Guerra, P. Ventosilla. London, it send to publish.

• 2 Publication will have made by the year 2007, about the Western blot, Elisa and Dot-blot.

Obstacles

• The main obstacles that our group found was that 2004 and 2005 there were few acute patients.

• We have not training as planning.