1

AMITY INSTITUTE OF
BIOTECHNOLOGY

INDUSTRIAL TRAINING REPORT

JAWAHAR LAL NEHRU MEDICAL
COLLEGE(JLN)

SUBMITTED BY- MUDIT SINGH

COURSE- BSc(H)BIOTECHNOLOGY
2022-2025
ENROLLMENT NO.- A20204422002
2
 3

ACKNOWLEDGEMENT

I take immense pleasure in thanking Amity Institute of

Biotechnology for giving me the opportunity to carry
out this summer project work.
I express my indebtedness to H.O.D Prof. Vinay
Sharma (AIB), Dr. Manali Datta (coordinator ) for kind
support and allowance for completing my summer
training.
I sincerely express my thanks and gratitude to Dr.
VIJAYLATA RASTOGI (Microbiology department
head of JLN Medical College) for her guidance and
approval in this project.
My deepest thanks to the Professors of AIB who have
guided me and motivated me throughout the project.
Finally, I would like to express my gratitude to my
beloved parents for their blessings, my colleagues and
classmates for their help and wishes for the successful
completion of my project.

Mudit Singh
BSc(H) Biotechnology
 4

INDEX

S.NO. PARTICULARS
1 INTRODUCTION
2 HEPATITIS-B
3 HEPATITIS-C
4 HIV
4.1 ELISA METHOD
4.2 TRI-DOT METHOD
5 WIDAL TEST
6 SEROLOGY TEST
6.1 RF TEST
6.2 ASO TEST
7 IDSP LAB
8 CONCLUSION
 21

INTRODUCTION

Medical microbiology involves the identification of

microorganisms for the diagnosis of infectious diseases and
the assessment of likely response to specific therapeutic
interventions. Major categories of organisms include bacteria,
mycobacteria, fungi, viruses, and parasites.

Microbiological methods combined with clinical symptoms,

additional laboratory tests, and imaging techniques are used in
combination to distinguish a true disease-associated infection
from colonization with normal flora or other conditions, such
as malignancies, inflammatory disorders, or autoimmune
disorders, all of which have unique therapies and prognoses
for the patient.

Diagnostic Medical Microbiology is concerned with the

etiologic diagnosis of infection.

The job of the clinical microbiology laboratory include –

1. To test specimens from patients for microorganisms that
are, or may be, a cause of illness.
2. To provide information about the in vitro activity of
antimicrobial drugs against the microorganisms
identified.
3. Confirming a clinical diagnosis of infectious disease with
a bacterial etiology.
4. To advice the physician as well as process specimens.
 21

HEPATITIS-B

Most common agent causing acute hepatitis is virus. Of these

Hepatitis B virus leads to severe forms of disease like liver disease,
including liver cancer and cirrhosis. This test is based on the
principle of immune-chromatography. The HBsAg Card consists of
four main regions i.e. the sample well, the conjugate pad, test
window, and the sink pad.
If HBsAg is present in the sample, the labeled monoclonal antibody
binds to it and moves upward.

1. Allow the HBsAg test pouch to attain room temperature.

2. Carefully remove the card from the pouch & place it on flat
clean surface.
3. Add 50µl or 2 drops of serum sample into the sample well
using the micro pipette or dropper.
4. Read results within 20 minutes.
 21

(HBsAg Rapid Card test)

HEPATITIS-C

Hepatitis C Virus (HCV) is a small, enveloped, positive-

sense, single-stranded RNA virus.
1. Add 3 drops of Buffer Solution to the center of the
device.
2. Hold the dropper vertically downwards and add 1
drop of patient's sample (50 µl serum or plasma) using
the sample dropper provided. (use a separate sample
dropper for each specimen to be tested).
3. Add 5 drops of Buffer Solution.
 21

4. Add 2 drops of Protein- A Conjugate.

5. Add 5 drops of Buffer Solution.
6. Read result immediately and discard the device
considering it to be potentially infectious.

(HCV Rapid Card Test)

1. Appearance of two dots, one at the control region

“C” & other at the test region “T1” indicates that the
sample is REACTIVE for antibodies to HCV.
2. Appearance of two dots, one at the control region
“C” & other at the test region “T2” indicates that the
sample is REACTIVE for antibodies to HCV.
3. Appearance of all the three dots, one each at “C”
 21

“T1” & “T2” region indicates that the specimen is

REACTIVE for antibodies to HCV.

HIV
Laboratory work in the HIV section of microbiology often
involves serological testing to detect the presence of HIV
antibodies or antigens.

1. Meriscreen (ELISA Method)

• Principle: Meriscreen is based on the Enzyme-Linked
Immunosorbent Assay (ELISA) method. It is used for the
qualitative detection of antibodies to HIV-1 and HIV-2 in
human serum or plasma.
• Procedure:
1. A sample of the patient's serum or plasma is added
to wells coated with HIV antigens.
2. If HIV antibodies are present, they bind to the
antigens.
3. A secondary antibody conjugated with an enzyme is
added, which binds to the HIV antibodies.
4. A substrate is then added, which reacts with the
enzyme to produce a color change.
5. The intensity of the color is measured, which
correlates with the presence of HIV antibodies.
 21

•Interpretation: A color change indicates a positive

result, suggesting the presence of HIV antibodies.

(ELISA Card)

(ELISA Card Marked with patient’s No.)

 21

(ELISA Wells)

(ELISA Card In Wells)

Tri-Dot (Rapid Test)

• Principle: The Tri-Dot test is another rapid test that
utilizes a dot-immunoassay technique for the detection of
HIV antibodies.
 21

• Procedure:
1. A drop of blood, serum, or plasma is added to a small
test strip or membrane.
2. HIV antigens are immobilized on the membrane in a
dot format.
3. If the sample contains HIV antibodies, they bind to
the antigens.
4. A secondary antibody conjugated with a dye is added,
which binds to the HIV antibodies.
5. A visible dot appears on the membrane, indicating a
positive result.

• Interpretation: The presence of a colored dot indicates

a positive test result for HIV antibodies.
 21

(HIV Rapid Card)

WIDAL TEST
A Widal test meaning is a sersology blood test that helps
detect typhoid or enteric fever in the body.
The main Widal test principle is that if a particular antibody is
present in the serum, it will react with a specific antigen and
show visible clumping on the test card.

 Collect a blood sample from the patient and let it clot.

 After the clot has formed, centrifuge the sample to
separate the serum from the cells.
 Place a drop of the serum on a clean glass slide and label
it with the patient's name and the date.
 21

 Add a drop of antigen solution (containing killed

Salmonella typhi and Salmonella paratyphi) to the serum
drop.
 Mix the two drops using a clean toothpick or glass rod,
making sure they are thoroughly combined.

 Rotate the slide slowly for 4 minutes to allow

agglutination to occur. Agglutination is the clumping of
bacteria caused by the interaction between the antibodies
in the serum and the antigens in the solution.
 Examine the slide under a microscope at 40x
magnification to look for agglutination. The clumping of
the bacteria indicates a positive result.
 21

(WIDAL Test Card)

SEROLOGY TEST

1. RF TEST- A rheumatoid factor (RF) test looks for

 21

rheumatoid factor (RF) in a sample of your blood. The

test looks for rheumatoid factors, which are proteins
produced by the immune system that can attack healthy
cells and tissues. The test can help diagnose autoimmune
disorders, such as rheumatoid arthritis (RA), as well as
chronic infections and certain types of cancer.
 Place one drop of the patient’s serum on a test card.
 Add a drop of RF latex reagent to the serum and mix
gently.
 Observe for agglutination (clumping) within 2 minutes.

 Positive Result: Visible agglutination indicates the

presence of RF.
 Negative Result: No agglutination means RF is either
absent or below detectable levels.
 21

(RF Latex Reagent)

2. ASO TEST- The Antistreptolysin O (ASO) test measures

the level of antibodies produced against streptolysin O, a
toxin released by group A Streptococcus bacteria. This
test helps in diagnosing a recent streptococcal infection,
which can lead to conditions like rheumatic fever.

 Place a drop of the patient’s serum on a test slide.

 Add a drop of the ASO latex reagent and mix gently.
 Observe for agglutination (clumping) within 2 minutes.
 21

 Positive Result: Visible agglutination indicates the

presence of ASO antibodies.
 Negative Result: No agglutination indicates that ASO
antibodies are either absent or below detectable levels.

(ASO Latex Reagent)

IDSP LAB
Quantitative Polymerase Chain Reaction (qPCR), also known
as real-time PCR, is a powerful diagnostic tool used to detect
and quantify the genetic material of pathogens in clinical
samples. This technology is especially valuable in detecting
infectious diseases such as hepatitis and dengue, where early
and precise detection is critical for treatment and control.
Given its high sensitivity, qPCR can detect even low levels of
viral DNA or RNA in blood samples, providing a reliable
method for early diagnosis.
 21

Principle of qPCR in Disease Detection - qPCR is an

advanced form of PCR that enables the amplification and
quantification of nucleic acids in real-time. In disease
detection, qPCR targets pathogen-specific genetic sequences
in blood samples, allowing for accurate identification and
quantification of the infectious agent. The amplification
process is monitored through fluorescent dyes or probes that
bind to the pathogen's DNA or RNA, emitting signals that
increase proportionally with the quantity of amplified product.

Application of qPCR in Detecting Hepatitis - Hepatitis

viruses, such as HBV (Hepatitis B Virus) and HCV (Hepatitis
C Virus), have unique genetic sequences that qPCR can target
for diagnosis. Blood samples are typically processed to isolate
viral RNA, which is then reverse-transcribed into
complementary DNA (cDNA). Specific primers and probes
targeting the viral genome are used to amplify and detect viral
RNA, allowing clinicians to determine viral load—a critical
factor for assessing disease severity and treatment response.
 21

(Truenat Cartridge)

(Micro Chip)
 21

Advantages of qPCR for Disease Diagnosis - High

Sensitivity and Specificity: qPCR’s ability to detect low levels
of viral nucleic acids in blood samples makes it highly
effective for early diagnosis.
Quantification: qPCR provides quantitative results, allowing
clinicians to monitor disease progression by measuring viral
load over time.
Speed: Compared to traditional diagnostic methods, qPCR
delivers results quickly, which is crucial for timely
intervention in infectious diseases.
Automation and High Throughput: qPCR systems can process
multiple samples simultaneously, making it suitable for large-
scale diagnostic settings

CONCLUSION-
My summer internship provided valuable hands-on-experience enhancing my
knowledge in medical microbiology. The exposure to real world challenges,
collaboration with diverse teams, and the guidance from mentors have
significantly contributed to my professional growth. I was deeply grateful for
this opportunity and look forward to applying these learnings in my future
endeavours.

end of report
 21

end of report