1

A REPORT ON

Rapid immunochromatography test for

HCV in human serum/plasma

BY

Name of Student: ID No:

Atharva Tandon
2016B1A80949P

Discipline:

M.Sc.(Hons) Biological Sciences with B.E. (Hons) Electronics

and Instrumentation

AT

QUALPRO DIAGNOSTICS, GOA

A Practise School-I station of

BIRLA INSTITUTE OF TECHNOLOGY & SCIENCE, PILANI

(JUNE, 2018)
 2

A REPORT ON

Rapid immunochromatography test for

HCV in human serum/plasma

BY

Name of Student: ID No:

Atharva Tandon
2016B1A80949P

Discipline:

M.Sc.(Hons) Biological Sciences with B.E. (Hons) Electronics

and Instrumentation

Prepared in partial fulfilment of the

Practise School-I course No.

BITS F221/BITS F231/BITS F241

AT
Qualpro Diagnostics, Goa

A Practise School-I station of

BIRLA INSTITUTE OF TECHNOLOGY & SCIENCE, PILANI
(JUNE, 2018)
BIRLA INSTITUTE OF TECHNOLOGY AND SCIENCE
 3

ACKNOWLEDGEMENT:

I would like to thank Mr. K. Sharma for making us comfortable

with the Qualpro work environment and encouraging us to learn
more about the diagnostic market. I would also like to thank Dr.
Sonali (Research and Development) who helped us a lot in
analysing the business model of the company. I would like to
thank the workers and employees whose expertise led to a great
deal of exposure. I would also like to thank my fellow group
members Surabhi and Lakshita, without whom this project could
not have been completed. Finally, I would like to express my
sincere gratitude to our PS instructor, Prof. Ravi Prasad Aduri,
for encouraging us to analyse the scientific principles as well as
the business front of the organisation and giving us a lot of
freedom.
 4

PILANI (RAJASTHAN)
Practise School Division

Station : Qualpro Diagnostics Centre:

Goa

Duration: From: 22nd May 2018 To : 14th July

2018

Date of Submission:13th July 2018

Title of the Project: Rapid immunochromatography test for HCV

in human serum/plasma

ID No. 2016B1A80949P Name: Atharva

Tandon Discipline : M.Sc.(Hons)
Biological Sciences with B.E. (Hons) Electronics and
Instrumentation

Name of expert: Mr. Kadam, Dr. Sonali

Designation: Manager and Research lead respectively

Name of the PS Faculty: Prof. Ravi Prasad Aduri

Key Words: Immunochemistry Market, SWOT analysis, Gold

coated membrane, Compound Annual Growth Rate, etc.

Project Area: Affordable Healthcare Technology

Abstract : Globally, an estimated 71 million people have
chronic hepatitis C infection, HIV continues to be a major global
public health issue, having claimed more than 35 million lives so
 5

far. In 2016, 1.0 million people died from HIV-related causes

globally. The widespread existence of these deadly diseases
prompts us to dwell more into the diagnostic sector. India, with
its very young demographic, is prone to these kind of threats.
Hence, understanding about the diagnostic kits and existing
technology becomes crucial in tackling this threat. The aim of
this report is to analyze the Indian diagnostic market as well as
understand the existing technology available for diagnosis and
suggest improvements. I have been assigned to study the Rapid
Diagnostic Test product for Hepatitis C Virus designed by
Qualpro Diagnostics. The product name is Flaviscreen HCV Plus.
The methodology used to analyse the product has two aspects.
First, is the scientific principle of the product and the second is
the cost analysis approach. The cost analysis would entail the
comparison of this product with its competitor. I have carried
out a detailed research about the product and provided some
solutions to improve the technology and suggest alternate
technology. The report also entails of a detailed SWOT analysis
which is based on observations

.
 6

TABLE OF CONTENTS:

i. Acknowledgement
ii. Abstract
iii. Introduction

1. Diagnostic Market
1.1 In Vitro Diagnostics
1.2 Immunochemistry Market
1.3 Key Players
2. Tulip Diagnostics
2.1 Brief Overview of the Organisation
2.2 National Sales
2.3 Distribution
2.4 International Sales
2.5 Merger with PerkinElmer
3. Quality Assessment Mechanism for Exports
4. Qualpro Diagnostics and Analysis of Flaviscreen HCV Plus:
4.1 Qualpro Diagnostics
4.2 Flaviscreen HCV Plus
 7

4.3 Other Products in the Market

5. Improvement in existing technology and alternate methods
5.1 Dried Blood Spots Along With RDTS:
5.2 Test Using Oral Fluids
5.3 Combination Integrated Multi Disease Test
5.4 Use Of Mobile Application to Enhance Connectivity
5.5 Alternate Technology
6. SWOT Analysis
7. Conclusion
8. References
 8

INTRODUCTION:
Globally, an estimated 71 million people have chronic hepatitis
C infection, HIV continues to be a major global public health
issue, having claimed more than 35 million lives so far. In 2016,
1.0 million people died from HIV-related causes globally. The
widespread existence of these deadly diseases prompts us to
dwell more into the diagnostic sector. India, with its very young
demographic, is prone to these kind of threats. Hence,
understanding about the diagnostic kits and existing technology
becomes crucial in tackling this threat. The aim of this report is
to analyze the Indian diagnostic market as well as understand
the existing technology available for diagnosis and suggest
improvements. I have been assigned to study the Rapid
Diagnostic Test product for Hepatitis C Virus designed by
Qualpro Diagnostics. The product name is Flaviscreen HCV Plus.
The methodology used to analyse the product has two aspects.
First, is the scientific principle of the product and the second is
the cost analysis approach. The cost analysis would entail the
comparison of this product with its competitor. I have carried
out a detailed research about the product and provided some
solutions to improve the technology and suggest alternate
technology. The report also entails of a detailed SWOT analysis
which is based on observations. Our team has tried a lot to make
the analysis as extensive as possible but there were certain
areas where we could not get the necessary data. The report
aims to give a holistic picture of the product, the organization
and aims to benefit people who wish to start a business or
understand more about the diagnostic industry.
 9

1. Diagnostic Market:
1.1 In Vitro Diagnostics:
In vitro diagnostics are tests done on samples such as blood or
tissue that have been taken from the human body. In vitro
diagnostics can detect diseases or other conditions, and can be
used to monitor a person’s overall health to help cure, treat, or
prevent diseases. In vitro diagnostics may also be used in
precision medicine to identify patients who are likely to benefit
from specific treatments or therapies. These in vitro diagnostics
can include next generation sequencing tests, which scan a
person’s DNA to detect genomic variations. In Vitro Diagnostics
is divided into 7 segments mainly Clinical/Biochemistry,
Immunochemistry, Hematology, Microbiology ,Urinalysis,
Coagulation and molecular diagnostics.
 10

1.2 Immunochemistry Market:

Since the company under study is leading the immunochemistry
segment of the market, it is important to understand this
particular area and analyse the overall revenue generated by
this sector.

• Immunoassays are biochemical tests that measure the

presence or concentration of a macromolecule or a small
molecule in a solution through the use of an antibody or
antigen Size & Growth
• Immunochemistry Market Immunochemistry is the second
largest segment in the Indian IVD market. Reagents
account for 90% of the market.
 11

• The global immunochemistry instruments and reagents

market is expected to reach $ 24.5 billion by 2019, growing
at a CAGR of 7.2 percent from 2014 to 2019.
• There is a rising focus on biomarker development across
the globe as they help in detection of various diseases in
their initial stage.

1.3 Key Players:

The table below shows the dominant players and the presence
of these companies in various segments of the in-vitro diagnostic
market. These include Roche, Abott Healthcare, Siemens,
Beckman, BD, Transasia.
 12

2. Tulip Diagnostics:
2.1 Brief Overview of the Organisation:
Tulip Diagnostics (P) Ltd was established in 1988 , comprises of
eight independent diagnostic units has emerged as the leading
manufacturer and marketer of in-vitro diagnostic reagents, kits
and instruments both nationally and internationally.
 13

While Tulip Diagnostics (P) Ltd., focuses on assay systems for

Immunohematology, Hematology, Rheumatology, Infectious
Diseases, Hemostasis and Instrumentation, its
Division Microxpress focuses on Dehydrated Culture Media,
Bases, Supplements, Reagent and Test kits, Sterile ready
prepared plates, Antimicrobial Susceptibility discs, Plant tissue
culture media and chemicals for Microbiology and
Mycobacteriology. Orchid Biomedical Systems, Qualpro
Diagnostics, Zephyr Biomedicals focus on Rapid membrane &
ELISA based immunodiagnostic platforms along with
chemiluminescence.

Viola Diagnostic Systems deals with Rapid immunoassays for

Parasitology range and infectious diseases.

Coral Clinical Systems focuses on Clinical Biochemistry and

Instrumentation while its division BioShields deals with High
Quality antiseptics and disinfectants for industrial, medical and
domestic use.

Tulip Diagnostics (P) Ltd. currently has seven manufacturing

sites in Goa and two in North India. The recent expansion of a
plant in Uttarakhand, which is specifically for in vitro
diagnostics, has been a major endeavor for Tulip during the
year. Additionally, the company will also be making substantial
facility additions in the times to come, depending on the
demand.

The group companies are owned, managed and run by highly

involved professionals and has continuously set new trends
through product development and improvements. A widely
acclaimed Biotech business magazine published by Cybermedia
publication, India, in its issue 7 Vol 5, has rated Tulip Group of
Companies as the No 1 diagnostic company in India.
 14

In Goa, the three biomedical companies focus on different kits,

which are as follows-
Qualpro Diagnostics manufactures one step membrane and Elisa
based tests and kits for Fertility, Infectious diseases and
Parasitology.
The company has approximately 8200 sq.ft of manufacturing
area.

Zephyr Biomedicals manufactures one step membrane tests for

Fertility, Infectious diseases, Parasitology, Cancer and Cardiac
Markers.
The company has approximately 19900 sq.ft of manufacturing
area.

Orchid Biomedical Systems manufactures one step rapid

membrane based tests for Fertility, Infectious diseases and
Parasitology.
Orchid has manufacturing facilities covering approximately 8200
sq.ft. area.
 15

2.2 National Sales:

The Company’s national business is built around twelve branch
locations, nationwide with product flow all over the country
through a diverse and efficient distribution network that
guarantees product availability, maintenance of cool chain and
customer requirements. The Company has a professional sales
team of around 400 sales/service professionals headquartered
all over the country to carry forward its customer contact and
sales programme, with a customer base of over 20000 customers
and 350 distributors.

2.3 Distribution:
 16

The company's products are distributed nationally by about 350

distributors spread across the length and breadth of the country.
This ensures market penetration and easy availability of the
groups products.

2.4 International Sales:

The company products are distributed across approximately 87
countries in South Asia, South East Asia, Middle East, Europe,
Africa, Russia and Latin America. The group products are also
bought directly by various NGO's and international agencies like
the UNICEF and the WHO for their various Health Programs. The
international sales comprises of group's own branded sales, OEM
/ neutral label products and contract manufactured products for
various national and international companies.

Angola Bangladesh Albania

Botswana Cambodia Austria
Burkina Faso China Belgium
Cameroon Indonesia Bulgaria
Egypt Iran Denmark
Ethiopia Japan France
Ghana Laos Germany
Ivory Coast Malaysia Greece
Kenya Mongolia Hungary
Liberia Myanmar Ireland
Madagascar Nepal Italy
Malawi Philippines Netherlands
Mali Srilanka Poland
 17

Mauritius Taiwan Portugal

Morocco Thailand Romania
Mozambique Vietnam Spain
Nairobi Switzerland
Nigeria Turkey
Sierra Leone UK
South Africa
Sudan
Tanzania
Togo
Tunisie
Uganda
Zambia
Zimbabwe

Russia & CIS Middle-East South America

Kazhakastan Jordan Brazil

Russia Lebanon Chile
Turkmenistan Muscat Colombia
Ukraine Qatar Guyana
Rep. of Azerbaijan Ecuador
Syria El Sal
UAE Mexico
Yemen Peru

Oceania

Austrailia
Papua New
Guinea
Solomon Islands
 18

Venezuela

2.5 Merger with PerkinElmer:

Perkin Elmer, a global leader committed to innovating for a

healthier world, has entered into a definitive agreement to
acquire Tulip Diagnostics Private Ltd. This helps Tulip group of
companies to get more funding for Research and Development.

3.Quality Assessment Mechanism for

Exports:
While analysing the company, it was imperative to know how
does the export of the products work. Since, we are dealing
with diagnostic kits, it is very important for an organisation to
maintain the highest standards of quality before introducing a
product in the foreign market. This process of determining the
sensitivity and sensitivity could be explained by three crucial
steps:
 19

1. Internal Validation: If a company wants to launch the

product internationally the first step is internal checking of the
product. This is done by the quality assessment programs of the
company itself.
2.External Validation: The second step is the external
validation which is done by specific organisations like NIBSC,
ICGEB,etc.
3. Re- Verification: Before launching the product the final step
is verification by a foreign agency.

4.Qualpro Diagnostics and Analysis of

Flaviscreen HCV Plus:
4.1 Qualpro Diagnostics:
Qualpro Diagnostics manufactures one step membrane and Elisa
based tests and kits for Fertility, Infectious diseases and
Parasitology.
 20

The company has approximately 8200 sq.ft of manufacturing

area.
The kits manufactured under the infectious diagnostic category
range are given below:
 21

The kits are passed through three different departments

Manufacturing, Quality Control and Packaging.
In Manufacturing department all the parts of the kits are
manufactured individually. For different types of kits there is a
separate department. The parts of the kits are made with
utmost care and the same part are of same size.
In Quality Control the parts of the kits are checked at various
levels. Each part is checked individually as well as after the
whole kit is completed. The efficiency should be 100% otherwise
the whole batch is wasted. The storage conditions during the
quality control should be static. The temperature should be
maintained.
The last part that comes is Packaging. Once the kit is tested
properly and all the results are true positive then the kit is
packed finally and sealed. The batch is dispatched and ready to
be exported.

4.2 Flaviscreen HCV Plus:

 22

INTRODUCTION:
QUALPRO HCV is a rapid, third generation two-site sandwich
immunoassay for the detection of total antibodies specific to
Hepatitis C virus (HCV in human serum / plasma). The test
employs a genotype cross-reactive multi-epitope recombinant
antigen derived from the Core, NS3, NS4 and NS5 regions of
multiple HCV genotypes. The double antigen sandwich system
ensures detection of all anti-HCV antibody isotopes (viz. IgG,
IgM, IgA etc.) to all major HCV genotypes.
SOME FACTS:
• Sensitivity:100%
• Specificity: 99.6%
• Number of Tests: 50
• Cost: Rs. 1500 per kit.

Rapid immunochromatography test for HCV in human

serum/plasma
 23

PRINCIPLE:
QUALPRO HCV utilizes the principle of agglutination of
antibodies / antisera with respective antigen in
immunochromatography format along with use of nano gold
particles as agglutination revealing agent. The conjugate pad
contains two components - a multi-epitope HCV recombinant
antigen conjugated to colloidal gold and rabbit globulin
conjugated to colloidal gold. As the test specimen flows through
the membrane test assembly the HCV recombinant antigen-
colloidal gold conjugate complexes with the anti-HCV antibodies
in the specimen and travels on the membrane due to capillary
action along with the rabbit globulin - colloidal gold conjugate.
This complex moves further on the membrane to the test region
(T) where it is immobilized by another multi-epitope HCV
recombinant antigen coated on the membrane leading to
formation of a pink to pink-purple coloured band. The absence
of this coloured band in the test region indicates a negative test
result. The unreacted conjugate and unbound complex, if any,
along with rabbit globulin- gold conjugate.
moves further on the membrane and are subsequently
immobilized by the Agglutinating sera for rabbit globulin coated
on the membrane at the control region (C), forming a pink to
pink-purple coloured band. This control band acts as a
procedural control and serves to validate the results.
 24

PROCEDURE:
1.The sealed aluminum foil pouch of QUALPRO HCV membrane
test assembly is brought to room temperature.
2. Pouch is teared along the "notch".
3. The membrane test assembly is removed. Once opened, the
membrane test assembly must be used immediately.
4. The membrane test assembly is labelled with specimen
identity.
5. The membrane test assembly is placed on a flat horizontal
surface.
6. Carefully dispense 10 µl of serum / plasma into the specimen
port “A”.
7. Three drops of sample running buffer is added into port “B”.
8. Observe the development of visible colored band at Test
region.
9. Positive result may be observed within 15-20 minutes.
10. The test should be considered invalid if the control band “C”
does not appear. The test is also invalid if neither the control
nor the test bands appear.
 25

REAGENTS AND MATERIAL SUPPLIED:

QUALPRO HCV kit has following components:

1. Membrane test assembly:

Stripped with multi-epitope HCV recombinant antigen and

Agglutinating sera for rabbit globulin along with HCV specific
antigens and rabbit globulin gold conjugate. Each membrane
test assembly is individually pouched along with single-use
desiccant.

2.Sample Running Buffer: Buffer containing surfactant and

preservatives.
3. Disposable Plastic Sample Applicators.

STORAGE AND STABILITY:

QUALPRO HCV is stable to the expiry date mentioned on the

label when stored at 4-30°C. Once the pouch is opened, the
device must be used immediately.

MATERIALS REQUIRED BUT NOT

PROVIDED:

1. Disinfectant
2. Disposable gloves
3. Biohazard waste container
4. Micropipette
 26

SAMPLE COLLECTION
1. QUALPRO HCV uses human serum / plasma as specimen.
2. No special preparation of the patient is necessary prior to
specimen collection by approved techniques.
3. Preferably use fresh sample. However, specimen may be
stored refrigerated (2-8°C) for short duration. For long storage,
freeze at -20°C or below.

INTERPRETATION OF RESULTS:

PERFORMANCE CHARACTERISTICS:

PPPPP
4.3 Other Products in the Market:
27
28
 29

5. Improvement in existing
technology and alternate methods:

5.1 Dried Blood Spots Along With RDTS:

 30

• The 2017 WHO Guidelines Development Group recognized

the benefits of DBS and in particular their potential to
facilitate greater access to testing and recommended
considering DBS for both serological and NAT HBV and HCV
technologies.
• Dried blood spot testing (DBS) is a form of bio sampling
where blood samples are blotted and dried on filter
paper.
• Combining RDTs and Dried blood samples provide better
accuracy and improved Point of Care Testing.

5.2 Test Using Oral Fluids:

• In the field of HIV, the use of oral HIV RDTs for community
based or home self-testing has increased access to testing
for those who are marginalised or reluctant to visit clinics
for testing due to stigma, and has been particularly
successful.
 31

• There is a possibility that the same increased access be

achieved in the case of HCV as well.
• For viral hepatitis, use of oral fluids is currently only
available with one quality approved HCV antibody RDT

OraQuick® HCV Rapid Antibody Test

5.3 Combination Integrated Multi

Disease Test:
• As HIV-infected persons continue to live longer due to
increased uptake of anti-retroviral therapies, liver disease
has emerged as a leading cause of morbidity and mortality
in persons co-infected with HBV or HCV.
• Combination integrated multi-disease blood or oral based
assays that allow for integrated testing of HIV, HBV and
HCV using a single sample would improve the efficiency of
screening programmes.
• A further innovation in HCV serology is a combination RDT
for the simultaneous detection of anti-HCV and core
 32

antigen, allowing for early detection of recent HCV

infection
• The HCV core Ag assay has shown to be as effective as NAT
in detecting HCV infection, and about 6–7 weeks earlier
than third generation anti-HCV assays

5.4 Use Of Mobile Application to

Enhance Connectivity:
• India is one of the leading consumer of smart phones. The
number of people coming online is increasing at a rapid
pace. Hence, it provides an opportunity to tap this hidden
potential in this sector.
• An accessible standardized database that enables
integration with laboratory information systems, electronic
health records, and health programme databases can allow
diagnostics to meet these broad-ranging need.
• Students and faculty at Stanford University have developed
a new smartphone device that can detect the hepatitis B
virus (HBV), as well as HIV, influenza, cardiovascular and
autoimmune disorders, cancer, and the hepatitis C virus
(HCV), via an on-the-go app
• The new device works by analyzing a drop of blood. The
machine identifies magnetically tagged biomarkers and
then displays a patient’s test results on their smartphone in
 33

less than 10 minutes. Results can then be sent back to

doctors via email or via the cloud on the device’s
supporting app.

5.5 Alternate Technology:

• Nucleic acid testing (NAT) technologies are highly accurate
in detecting the presence of viral nucleic acid (DNA or
RNA), and are used to confirm either the presence or
absence (qualitative) or level (quantitative) of viraemia to
guide treatment decisions and monitor treatment response,
and confirm either virological cure (HCV) or effective
suppression (HBV).
• NAT assays can now be performed on POC devices outside
of laboratory settings. Point-of-care testing or near patient
testing is defined as medical diagnostic testing at or near
the point of care—that is, at the time and place of patient
care

6. SWOT Analysis:
SWOT stands for: Strength, Weakness, Opportunity, Threat. A
SWOT analysis guides you to identify your organization’s
strengths and weaknesses (S-W), as well as broader
opportunities and threats (O-T). Developing a fuller awareness
of the situation helps with both strategic planning and decision-
making.
 34

SWOT Analysis of Tulip Group:

Strengths:
• Close proximity to a major port.
• Tremendous national and international sales.
• Wide Distributor Network
• Dominating the Indian Reagent market
• Good product line up.
• Merger with Perkin Elmer.
• Expansion in North India.
• Long established player.

Threats:
• Increased Competition.
• The recent curbing of plastic might lead to restrictions
in manufacturing and production.
• Lack of regulation. Parliament enacted the Clinical
Establishments Act, 2010, which prescribes minimum
standards for facilities and services for clinical
establishments, but this has to be to adopted by states
as health is a state subject.

Weakness

• Not adequate funds diverted to R&D.

• Not much presence in social media or digital marketing.

Opportunities

• Cost cutting and improved production by automation.

• Less transportation costs - GST
 35

• Could enter the thyroid market to increase dominance

in immunochemistry market.
• Chemiluminescent systems are gaining
momentum(CLIA).
• Growing number of insured lives.
• Changing demographics and prevalence of disease.
• Expansion of pathology diagnostic market to towns and
rural areas
• Social media to increase brand awareness.
• Increase in the number of hospitals and healthcare
facilities, development of healthcare infrastructure and
the advancement of medical equipment National Health
Protection Scheme (NHPS)
 36

7. Conclusion:

We have established the need for Rapid Diagnostic Test and

analysed the business model of one of these RDTs in report. The
conclusion of this report recognizes Tulip as a major company in
the Immunochemistry Market with a potential to grow more
after the merger with Perkin Elmer. It needs to focus more on
 37

Research and Development to produce more innovative

products. An mobile application based approach would be useful
in storing data with RDTs.

8. References:

• https://www.grandviewresearch.com/industry-analysis/in-vitro-
diagnostics-ivd-market

• https://www.prnewswire.com/news-releases/indian-diagnostic-
opportunities-2018-226133631.html

• https://www.hepmag.com/article/smartphone-hepatitis-hiv-
testing-26583-677428900

• https://www.ncbi.nlm.nih.gov/pubmed/23902574

• http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0074-
02762012000500016

• https://www.fda.gov/medicaldevices/productsandmedicalproced
ures/invitrodiagnostics/default.htm