Skip to main content
main-content

01.12.2012 | Methodology | Ausgabe 1/2012 Open Access

Malaria Journal 1/2012

Proteomic identification of host and parasite biomarkers in saliva from patients with uncomplicated Plasmodium falciparum malaria

Zeitschrift:
Malaria Journal > Ausgabe 1/2012
Autoren:
Honglei Huang, Mukram M Mackeen, Matthew Cook, Eniyou Oriero, Emily Locke, Marie L Thézénas, Benedikt M Kessler, Davis Nwakanma, Climent Casals-Pascual
Wichtige Hinweise

Electronic supplementary material

The online version of this article (doi:10.​1186/​1475-2875-11-178) contains supplementary material, which is available to authorized users.

Competing interest

All authors declare that they have no competing interest.

Authors’ contributions

HH, MM, MC, EO, MLT carried out the proteomic studies and drafted the manuscript. BMK, DN and CCP participated in the design of the study, performed the statistical analysis. MM, EO, DN, EL and CCP conceived the study, and participated in its design and coordination. All authors read and approved the final manuscript.

Abstract

Background

Malaria cases attributed to Plasmodium falciparum account for approximately 600,000 deaths yearly, mainly in African children. The gold standard method to diagnose malaria requires the visualization of the parasite in blood. The role of non-invasive diagnostic methods to diagnose malaria remains unclear.

Methods

A protocol was optimized to deplete highly abundant proteins from saliva to improve the dynamic range of the proteins identified and assess their suitability as candidate biomarkers of malaria infection. A starch-based amylase depletion strategy was used in combination with four different lectins to deplete glycoproteins (Concanavalin A and Aleuria aurantia for N-linked glycoproteins; jacalin and peanut agglutinin for O-linked glycoproteins). A proteomic analysis of depleted saliva samples was performed in 17 children with fever and a positive–malaria slide and compared with that of 17 malaria-negative children with fever.

Results

The proteomic signature of malaria-positive patients revealed a strong up-regulation of erythrocyte-derived and inflammatory proteins. Three P. falciparum proteins, PFL0480w, PF08_0054 and PFI0875w, were identified in malaria patients and not in controls. Aleuria aurantia and jacalin showed the best results for parasite protein identification.

Conclusions

This study shows that saliva is a suitable clinical specimen for biomarker discovery. Parasite proteins and several potential biomarkers were identified in patients with malaria but not in patients with other causes of fever. The diagnostic performance of these markers should be addressed prospectively.
Zusatzmaterial
Additional file 1: Lectin depletion using Concanavalin A (ConA) and optimization of starch depletion. The figure shows the optimization of ConA depletion and starch depletion. (Key: L = Ladder, E = Elute, FT = Flow Through, W1 = Wash 1, W2 = Wash 2, AR = Amylase-removed Control, Ctrl = Control sample of saliva). (DOCX 234 KB)
12936_2012_2119_MOESM1_ESM.docx
Additional file 2: Optimization of lectin depletion using AAL. The figure shows the optimization of saliva sample volume, assessing the effectiveness of 50 μg lectin depletion of 50, 100, 200 and 400 μl of amylase removed saliva sample. The figure shows the differential band patterns in the FT and E showing successful depletion strategies. (Key: L = Ladder, E = Elute, FT = Flow Through, W = Wash, AR = Amylase Removed Control, AR-E = Elute from the Starch used to remove amylase, Ctrl = Control sample of saliva). (DOCX 384 KB)
12936_2012_2119_MOESM2_ESM.docx
Additional file 3: Optimization of lectin depletion using jacalin. The figure shows the optimization of saliva sample volume, assessing the effectiveness of 50 μg lectin depletion of 50, 100, 200 and 400 μl of amylase removed saliva sample. The figure shows the differential band patterns in the FT and E showing successful depletion strategies. (Key: L = Ladder, E = Elute, FT = Flow Through, W = Wash, SDS = Final Elute using SDS loading buffer, AR = Amylase Removed Control, Ctrl = Control sample of saliva). (DOCX 340 KB)
12936_2012_2119_MOESM3_ESM.docx
Additional file 4: Optimization of lectin depletion using peanut agglutinin (PNA). The figure shows the optimization of saliva sample volume, assessing the effectiveness of 50 μg lectin depletion of 50, 100, 200 and 400 μl of amylase removed saliva sample. The figure shows the differential band patterns in the FT and E showing successful depletion strategies. (Key: L = Ladder, E = Elute, FT = Flow Through, W = Wash, SDS = Final Elute using SDS loading buffer, AR = Amylase Removed Control, Ctrl = Control sample of saliva). (DOCX 287 KB)
12936_2012_2119_MOESM4_ESM.docx
Authors’ original file for figure 1
12936_2012_2119_MOESM5_ESM.pdf
Authors’ original file for figure 2
12936_2012_2119_MOESM6_ESM.pdf
Authors’ original file for figure 3
12936_2012_2119_MOESM7_ESM.pdf
Literatur
Über diesen Artikel

Weitere Artikel der Ausgabe 1/2012

Malaria Journal 1/2012 Zur Ausgabe

Neu im Fachgebiet Innere Medizin

Meistgelesene Bücher aus der Inneren Medizin

2017 | Buch

Rheumatologie aus der Praxis

Entzündliche Gelenkerkrankungen – mit Fallbeispielen

Dieses Fachbuch macht mit den wichtigsten chronisch entzündlichen Gelenk- und Wirbelsäulenerkrankungen vertraut. Anhand von über 40 instruktiven Fallbeispielen werden anschaulich diagnostisches Vorgehen, therapeutisches Ansprechen und der Verlauf …

Herausgeber:
Rudolf Puchner

2016 | Buch

Ambulant erworbene Pneumonie

Was, wann, warum – Dieses Buch bietet differenzierte Diagnostik und Therapie der ambulant erworbenen Pneumonie zur sofortigen sicheren Anwendung. Entsprechend der neuesten Studien und Leitlinien aller wichtigen Fachgesellschaften.

Herausgeber:
Santiago Ewig

Mail Icon II Newsletter

Bestellen Sie unseren kostenlosen Newsletter Update Innere Medizin und bleiben Sie gut informiert – ganz bequem per eMail.

© Springer Medizin 

Bildnachweise