XIV IAMM AP CHAPTER ANNUAL STATE CONFERENCE JANUARY 2011

7TH -9TH

PG SYMPOSIUM : BIOMARKERS IN INFECTIOUS DISEASES

Guide: Dr. K. Saileela Professor, KIMS, Narketpally Dr. K.Pavani PG Microbiology

Definition:
A specific physical trait used as an indicator of normal biological process, pathogenic process or pharmacological response to therapeutic intervention.

It is as if the parasite left his own signature in the infected person

Biomarkers can be either pathogen or host derived. The pathogen derived biomarkers are highly specific, while host derived may be specific or nonspecific.

Biomarkers are broadly classified into: Proteins Nucleic acids LMW Metabolites

 Nucleic acid based biomarkers(pathogen derived) are

extremely powerful both in terms of accuracy of their identification and limit of detection that can be achieved using PCR amplification. It fails to provide information to assess a particular disease state .

Metabonomics refers to quantitative measurement of metabolic

responses within host-parasite system in response to infection or drug treatment It is restricted by numerous levels of regulation including life cycle specific gene expression, subcellular protein localization, enzyme concentration, post translational regulation, metabolite transport and substrate, product & cofactor abundance. These may be products of other microflora or involved in inflammation

Proteomics:
Since proteins are the main catalysts, structural elements,

signalling messengers, and molecular machines of biological tissues, proteomic studies are able to provide substantial clinical relevance.
Proteins can be utilized as biomarkers for tissues, cell

types, developmental stages, and disease states as well as potential targets for drug discovery and interventional approaches.
Recent advances in this area are attributable largely to the

introduction of mass spectrometry platforms capable of screening complex biological fluids for individual protein and peptide biomarkers.

Biomarkers for parasitic diseases :

PARASITIC DISEASE BIO MARKER

Chagas disease MIP1 alpha, C3a anaphylatoxin, and unusually truncated : forms of fibronectin, apolipoprotein A1 (ApoA1), and C3 Malaria Kalaazar C5a, Neopterin and Calcitonin Lepp12 antigen

Schistosomiasis Neutrophil gelatinase associated lipocalin (NGAL ) Cysticercus clusterin, lecithin-cholesterol acyltransferase, vitronectin, haptoglobin and apolipoprotein A-I.

Biomarkers for parasitic diseases :

PARASITIC DISEASE BIO MARKER

Onchocerciasis ESP

Toxoplasma gondi F. hepatica

Chorismate synthetase, nucleotide triphosphatases, Major surface antigen. Transferrin, Apolipoprotein A-IV

 N.Deckers, P. Dorny etal(Belgeum) : Porcine cysticercosis

Objective of study was to identify novel biomarkers in serum of experimentally infected pigs using SELDI-TOF MS
Momar Ndao, Terry W. Spithhill etal (Canada) :Chagas disease

Sera of patients, uninfected control & subjects with other parasitic diseases was compared using protein chip array MS.
Rene te Witt, Marlies E Van Wolfswinkel et al(Netherlandes):

Plasmodium falciparum Diagnostic accuracy of plasma soluble TREM-1, Neopterin & Procalcitonin as biomarkers in P. falciparum were evaluated in non P. falciparum, uncomplicated P. falciparum & severe P. falciparum malaria

 Si-Hwan Choi, Tae Yun Kim etal (Korea) : Toxoplasma gondi

Proteomic characteristics of T. gondi KI-1 tachyzoites from korean patients was compared with well known virulent RH strain. Protein composition of KI-1 tachyzoites & RH strain is similar but differential protein expression was involved in virulence.
Yulan Wang, Elaine Holmes etal: Schistosoma mansoni

S. mansoni cercaria infected mice urine samples collected 50 days post infection were compared with control urine samples from uninfected mice. Decreased TCA intermediates & disturbed aminoacid metabolism was detected.
Judith R. Denery, Ashlee A.K. Nunes etal(USA): O. volvulus

Analysis of African samples set revealed set of 14 biomarkers that discriminate O. volvulus positive & negative individuals. The biomarker set could also distinguish individuals with worms of compromised viability from those with active infection.

Techniques used for the expression analysis of proteins are:

Two-dimensional (2D) gel electrophoresis Matrix-assisted laser desorption ionization time-of-flight

mass spectrometry (MALDI-TOF MS) Surface-enhanced laser desorption ionization time of flight mass spectrometry (SELDI-TOF MS) Liquid Chromatography combined with MS (LCMS) Isotope-coded affinity tags (ICAT) Isotope tags for relative and absolute quantification (iTRAQ)

PRINCIPAL OF MASS SPECTROSCOPY

MALDI target plate:

SELDI-TOF Mass Spectrometer

PRINCIPAL OF SELDI-TOF CHIP ASSAYS

Advantages: Diagnosis of infection Distinguishes between past, latent, acute & reactivated infection. Determining risk of developing disease Determining stage of disease Determining response to therapy or prognosis. Measuring relevant biomarker & comparing them with reference amount or pattern of biomarkers associated with particular risk level/stage.

Disadvantages: Protocol dependence Identification limited to strains covered within database Taxonomical classification is difficult Operator/ Instrument variability Software limitations Strain identification difficult

References: 1. Mc Donald, W.H. & Yates, J.R. 3rd . Shot gun proteomics & biomarker discovery. Dis. Markers, 2002, 18, 99-105. 2. Ndao, Momar etal. Is SELDI-TOF a valid tool for diagnostic biomarker. Trends in Parasitology, 26(12), p.561, Dec 2010. 3. Wang, Yulan etal. Advances in metabolic profiling of experimental Nematode & Trematode infections. Advances in Parasitology, 73, p.373, Jan 2010. 4. Moma Ndao, Terry W,Spithill et al.Identification of novel diagnostic serum biomarkers for chagas disease in asymptomatic subjects by mass spectrometric profiling.Journal of Clinical Microbiology,April 2010,pg 11391149.