WORLD JOURNAL OF PHARMACY AND PHARMACEUTICAL SCIENCES

Ravishankar et al. World Journal of Pharmacy and Pharmaceutical Sciences

SJIF Impact Factor 5.210

Volume 4, Issue 12, 344-349 Review Article ISSN 2278 4357

OVER VIEW ON DEVELOPMENT AND APPLICATIONS OF

IMMUNOCHROMATOGRAPHY

B. Ravishankar1*, R. Rajesh2, E. Venkatanagaraju3, G. Divakar4, T. Shreeshail5,

Y. Nehalatha Reddy6

1,2
Department of Pharmaceutical Analysis, Acharya & BM Reddy College of Pharmacy,
Soldevanahalli, Hesaraghatta, Bengaluru-560107, Karnataka, India.
3, 4
Department of Pharmaceutical Biotechnology and Microbiology, Acharya & BM Reddy
College of Pharmacy, Soldevanahalli, Hesaraghatta, Bengaluru-560107, Karnataka, India.
5
Department of Pharmaceutics, Acharya & BM Reddy College of Pharmacy, Soldevanahalli,
Hesaraghatta, Bengaluru-560107, Karnataka, India.
6
Department of Pharmaceutics, Gokaraju Rangaraju College of Pharmacy, Nizampet Road,
Bachupally, Hyderabad - 500 090, Telangana, India.

Article Received on ABSTRACT

02 Oct 2015,
This review article gives an overview of latest research involving the
Revised on 23 Oct 2015, use of Immunochromatography for qualitative and quantitative
Accepted on 12 Nov 2015
analysis in different areas. The importance and versatility of detection
formats make these strips an ideal in various applications of Clinical
*Correspondence for
Author diagnosis, biological infectious agents and chemical contaminants.
B. Ravishankar Outline of review discuss the principle, developments that can be done
Department of with Immunochromatography and its applications. This overview can
Pharmaceutical Analysis,
provide future challenges and new strategies that can be done in the
Acharya & BM Reddy
area of immune chromatography.
College of Pharmacy,
Soldevanahalli,
KEYWORDS: Immunochromatography, clinical diagnosis, biological
Hesaraghatta, Bengaluru-
560107, Karnataka, India. infectious agents, chemical contaminants.

INTRODUCTION
Immunochromatography is one of the most important and effective technique in the detection
of virus and bacteria.[1] It plays an important role in the diagnosis. These assays are also
known as lateral flow test or simply strip test which are the devices intended to detect the
target analyte in sample without the need for specialized and costly equipment. They are the

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Ravishankar et al. World Journal of Pharmacy and Pharmaceutical Sciences

logical extension of the technology used in latex agglutination tests, the 1 st of which was
developed in 1956 by Singer and Plotz. The principle is based on dye labelled antibody
specific for target analyte which is present on the lower end of nitrocellulose strip or in the
plastic well along with the strip.[2] The antibody which is specific for target antigen is also
bound to the strip in a thin test line and antibody antigen specific for labelled antibody bound
to control line [Fig: 1]. So when the sample and buffer are placed on strip or in a well-mixed
with labelled antibody to draw across the lines of bound antibody. This gives the
identification of antigen present or absent. If the antigen is present then some of the labelled
antibody will be trapped on the test line and the excess labelled antibodies are trapped on the
control line [Fig: 2]. Currently, the principles governing this test are being extended and
forwarded to allow for some exciting new possibilities for future tests. The benefits of
Immunochromatography tests include: (i) They can be of users friendly format (ii) Very
short time to get test result (iii) Possess long term stability over a wide range of climates.(iv)
Relatively inexpensive to make.[1,2]

Fig.1: Nitrocellulose strip.

Fig.2: Identification for the presence and absence of antigens.

DEVELOPMENT POSSIBILITIES

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Ravishankar et al. World Journal of Pharmacy and Pharmaceutical Sciences

The possibility of creating a truly quantitative test can be done by using the same format of
lateral flow tests and dyeing the solid support with a fluorescent dye. The amount of antibody
bound at the capture line can be precisely quantified using a fluorimeter if the spectral
properties of the dyed microspheres to which the antibodies are conjugated are known. The
currently existing all lateral flow tests would provide all benefits to this and becomes
theoretically a truly quantitative assay. By placing multiple lines of captured antibodies on
the membrane, for an each different analyte, an individual can develop a single test for more
than one analytes. An exact or obvious application for this is to create drugs of abuse test
panel. Biosites Triage is based on this pattern. This principle diagnostically could be used
for panels of which all multiple analytes can be tested i.e, immune diseases, allergies or
multiple chemical sensitivity disorder. As the technology involved in preparing these tests
continues to be developing and progressing, it is possible to combine both of these ideas, and
to make a low-cost, rapid quantitative diagnostic assay for these multiple analytes.[3] Another
promising possibility of this technology is in the field of environmental sciences which
provides an opportunity to develop rapid & reliable tests that can be performed in the fields
of water pollution to plant disease. As these diagnostic tests must often be performed in harsh
environments lateral flow test format is unique for which, proper preparation, foil pouching
and no refrigeration or special handling is required. As scope in the field of molecular
genetics continues to expand rapidly, the focus in using a simple format for detecting various
genetic markers, DNA and RNA related disease infectious pathogens is increasing. The core
principle behind this type of test is the ability a ligand from solution to bind with a solid
support can be performed on genetic material as well as proteins, making this application of
the technology in this field almost a limitless. Immunochromatography with protein coated
microspheres (Proactive streptavidin) allows in optimizing direct attachment of antibody. In
this way, a series of tests can be developed rather quickly, using the same solid support,
membrane and housing etc.[4] Immunochromatography detection sensitivity is increased by
replacing gold colloidal platinum and newly developed silver amplification technology
amplifies particles more than 100 times quickly and increase sensitivity. This amplified
immunochromatographic system is expected to be more useful for specimen under the
condition liable to cause a false negative result.[5] The human immunodeficiency virus (HIV)
pandemic has become one of the greatest infectious disease threats to human health and
social stability that the world has ever faced. Every year around 6 lakhs new born children
will get infected with HIV worldwide. A pregnant women with HIV infection has an
approximately 30% chance of passing the virus to her born baby, prevention of parent to

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Ravishankar et al. World Journal of Pharmacy and Pharmaceutical Sciences

child transmission is a specific programme that provides a comprehensive family centred

spectrum of support and clinical services along with other public health initiatives to prevent
the transmission of HIV from parent to baby. So for this the sensitivity and specificity of a
Immunochromatography tests was 100% as compared to ELISA.[6]

APPLICATIONS
1. Immunochromatography along with google glass based for RDT reader capable of
qualitative and quantitative measurements of various lateral flow test and biomedical
diagnostic test.[7]
2. Immunochromatography along with fluorescent bead provides new strategies to
prevent the early stage transmission viruses in humans during both seasonal outbreak and
pandemics.[8, 9]
3. Immunochromatography is used for detection of human pluripotent stem cells
employing gold nano particles as a label and it was capable of detecting down to 10,000 cells
by visual inspection and 7000 cells by strip reader.
4. Used to detect clenbuterol a compound in the urine using fluorescent Nano silica and
visual detection limits.[10]
5. Used to determine ultra-small amounts of crustacean protein in processed foods which
can lead to allergic reaction.[11]
6. Used to quantify ractopamine in swine urine which was used as feed and can be toxic
to humans.
7. Used to identify nucleic acids by using recognition properties of molecular beacons
and optical properties of gold nano particles.[12]
8. Antibodies to phenolic glycolipid-1 of mycobacterium leprae were detected using
LFA for classification of leprosy patients and results showed good agreement with ELISA.[13]
9. Prostate specific antigen which is thought to be reliable marker for early diagnosis of
prostate cancer can be determined in human serum using gold nano particles as reported and
electrochemical detecting systems.
10. A simple sensitive and rapid visual detection of Hg+2 ions in aqueous solutions was
achieved by using gold nanoparticles in LFA for co-ordination events of Hg+2 between
thymine rich heparin oligonucleotide and digoxin labelled DNA probes which was
complimentary to a part of heparin oligonucleotide.[14]
11. Immunochromatography with use of immunogold conjugate helps in simultaneous
detection of carbofuran and triazophos in water samples.[15]

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12. LFA was used for detection of paraxon methyl using Fe3O4 aggregates as a label and
fluorescence step leader as detector.[16]

ACKNOWLEDGMENTS
The authors would like to thank the chairman Acharya Institutes Sri B.Premnath Reddy for
providing laboratory facilities and supporting this work.

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