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Bartonella (2003-2007).

P. Ventosilla, Sandra Palma,


Sandra Villar, Marcos Gonzales,
J. Chauca, H. Guerra,
C. Maguia.

OBJECTIVE
To development
immunological tests for
diagnosis, research and
transfer to the community
(2003-2007).

Colombia
Ecuador

Per

AMAZONAS

CAJAMARCA

PIURA

AMAZONAS

PIURA

PIURA

CAJAMARCA

CAJAMARCA

SAN MARTIN

AMAZONAS

AMAZONAS

AR CA
CAJAM

BARTONELLOSIS REPORTED BY
DEPARTMENT IN PERU
LORETO

SH
CA
AN

SAN MARTIN
LA LIBERTAD
ANCASH

ANCASH

HUANUCO

HUANUCO

HUANUCO

M
LI

ANCASH

LIMA

LIMA

LIMA

JUNIN

1995

CUSCO

CUSCO

AYACUCHO

1997

1999

2001

3258 cases
12 dead
12 departments

2005

2006

MINSA/OGE- 2006

Before DIGOS Support:

Birtles R, Fry N, Ventosilla P, Cceres A, Snchez 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).

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).

PA= Pooled acute phase sera


PV= Pooled chronic phase sera
PN= Pooled negative control sera

Results: Act 6.5.1.1. Western Blot using


individual Protein Antigens (Bands):
Table 1. Sensitivity and
specificity in the acute phase

Table 1. Sensitivity and


specificity in the chronic phase

M. W.
Band

Sensitivity

Specificity

M. W.
Band

Sensitivity

Specificity

65 kDa

76.3 %
(29/38)

87.5 %
(28/32)

65 kDa

86.1 %
(31/36)

87.5 %
(28/32)

12 kDa

42,1 %
(31/36)

100 %
(32/32)

12 kDa

88.8 %
(32/36)

100 %
(32/32)

26.3 %
(10/38)

100 %
(32/32)

13 kDa

41.6 %
(15/36)

100 %
(32/32)

13 kDa

Results: Act 6.5.1.2. ELISA


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 6.5.13.:


Standardization of DOT-BLOT
Sera: 1/50
Conjugate:1/300
Crude Antigen:
2mg

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 tutor
of 4 international students (Holland, USA,Germany
and 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 have
trained in clinical
and laboratory
diagnosis of
Bartonelosis about
115 persons from the
Health Minister and
non governmental
institutions.

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 ; Maguia 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; Maguia, Ciro; Best Ivan; Villar Sandra; Palma Sandra;
Henriquez Csar; 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. Gonzles, C. Prause, C. Maguia, P. Ventosilla. Two
immunological test to diagnostic carrions 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 Inmunolgico en Pacientes con
Bartonelosis Humana. E. Huarcaya, I. Best, J. Merello,
C. Maguia, P. Ventosilla. Aceptado para la publicacin
por Fundacin Instituto Hiplito Unnue, en Diagnstico.
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. Maguia, 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.

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