NAME: SAMRAT MUKHERJEE

YEAR: 3rd

SEMESTER: 5th

REGISTRATION NO: 15403319044

VENUE OF TRAINING: PEERLESS HOSPITEX HOSPITAL

AND B.K. ROY RESEARCH CENTRE LIMITED

DEPARTMRNT: MOLECULAR BIOLOGY LAB(RT-PCR)

TENURE : 8TH JULY,2021 TO 30TH OCTOBER,2021

ACKNOWLEDGEMENT

I would like to express my gratitude and appreciation to all those

who gave me the possibility of completing this report. A special thanks

to our principal Dr Sanjuckta Nandy; Mr Surajit Das, Head of

the Department, WBUT for giving me the permission to prepare

this report.

A special thanks to Mrs. Anindita Banerjee, HR in Peerless Hospital

for giving me opportunity to complete my training at Peerless

Hospitex Hospital and Research Centre Limited.

I also express my sincere gratitude to Mr Anupam Das, Senior

Manager (Quality Assuarance Department) At Peerless hospital,

Mr Ayan Kar (Executive in Lab Quality), Mr. Avishek Bera (Executive

in Lab Quality) for their co-operation, guide and support at the time

of training period.
DECLARATION
I hereby declare that the entire project report entitled “COVID-
19,RT-PCR’’ at Peerless Hospitex Hospital & Research Center
Limited is prepared by me during the academic year 2021 under
the guidance of Mr. Anupam Das Sir (Senior Manager, Quality
Assurance & Medical Administration). During the course of this
project, my guide both academic as well as company guide
helped me immensely.
I solemnly declare that whatever information I have used for
the completion of the project are true to the best of my
knowledge & the entire analysis and findings of the project
would be used for the academic purpose only.

…..……………………………….
Samrat Mukherjee
Bachelor of Hospital
Management 5th semester
Dinabandhu Andrews Institute of
Technology and Management
TO WHOM IT MAY CONCERN
This is to certify that. Mr Samrat Mukherjee, a student
of “Bachelor Of Hospital Management” from
Dinabandhu Andrews Institute Of Technolgy And
Management has undergone training in Quality
Assurance And Clinical Research Department of Peerless
Hospitex Hospital & Research Center Limited from 8th
July ,2021 to 30th October,[Link] has learned many
practical aspects of working in the department. I hope
this experience will help him in future. I wish him all
success in his future career.

Anupam Das
Senior Manager Quality Assurance (Internal Guide)
& Medical Administration
(External Guide)

–––––––––––––––––––––––––

HR Department
INDEX
TOPIC PAGE NO

INTRODUCTION OF PEERLESS HOSPITAL 6

COVID 19 7

ROLE OF MOLECULAR BIOLOGY LAB 8

SPECIMEN COLLECTION DETAILS 9

RT-PCR 10

SANITIZATION PROCESS IN LAB 14

STEPS FOR RT-PCR TESTING 16

JOBROLE DURING TRAINING 17

CONCLUSION 17
PEERLESS HOSPITAL

Peerless Hospital; located in Panchasayar area, Kolkata; offers the best treatment
to its patients, the clinic is well-equipped to offer the best treatment to patients. With
state-of-the-art infrastructure, equipment, dedicated doctors and well-trained staffs
are dedicated to assure to provide top-notch services.

Peerless Hospital is a renowned hospital in the Kolkata as well as for the neighbour
countries also. It is a 400 bedded [hospital as well as a research centre. Peerless
Hospitex Hospital &Research Centre limited is a multidisciplinary super-speciality
hospital. It is located within the screen and unpolluted natural surroundings near the
southern periphery of Kolkata. It is a referral centre of eastern India and nearby
countries. It has already become a trusted name in reliable, dependable and
advanced health care system for nearly twelve years. The hospital has most of the
major clinical and medicine specialties. The departments are fully equipped with the
latest advanced and state of the art diagnostic and therapeutic gadgets. The indoor
department has 400 beds in the forms of wards and cabins. Fifty-one beds are
equipped for providing intensive care.
COVID-19
COVID-19 is a disease caused by a new strain of coronavirus. “CO” stands for corona,
“VI” for virus and “D” for disease. Formerly this disease was referred to as “2019
novel coronavirus’ or 2019-nCoV’.

Corona viruses are a large family of viruses that are known to cause illness ranging
from the common cold to more severe diseases such as Middle East Respiratory
Syndrome (MERS) and Severe Acute Respiratory syndrome (SARS).

Symptoms of COVID-19: -
COVID19 affects different people in different ways. Most infected people will develop
mild to moderate illness and recover without hospitalization.

Most common symptoms: -

 Fever
 Dry cough
 Tiredness

Less common symptoms: -

 Aches and pains
 Sore throat
 Diarrhoea
 Conjunctivitis
 Headache
 Loss of taste or smell

On average it takes 5-6 days from when someone is infected with the virus for
symptoms to show, however it can take up to 14 days.
Corona Virus(COVID-19) Treatments: -
At home Corona virus Treatment: -

If your symptoms are mild enough that you can recover at home, you should:

 Rest. It can make you feel better and may speed your recovery.
 Stay home. Don’t go to work,schoolor public places
 Ask my doctor about over-the-counter medicines that may help, like
acetaminophen to lower my fever.

Time are cover from COVID-19

The strict home isolation for 7 days warning signs explained. Drink plenty of oral
fluids & Clinical follow up at 14th day & 28th day after discharge.

ROLE OF MOLECULAR BIOLOGY LAB

Molecular biology encompasses the structure and function of genetic material, its
transcription into RNA, and then translation into protein, as well as recently-
emerging regulatory processes, such as the roles of small RNA molecules (e.g.
miRNA, siRNA). DNA sequencing has undergone a recent revolution – while the
dependable Sanger method is still used, researchers have quickly taken advantage of
the faster “next-generation” sequencing methods. Amplifying DNA in the lab has
also become faster – PCR has evolved into the quicker “fast PCR,” as well as the
quantitative method of real-time PCR. Both DNA and RNA can be screened quickly
using microarray technology, which has grown in capacity to include tens of
thousands of samples per run. Genotyping organisms is efficiently accomplished
using mapping arrays on genotyping chips. For studying gene expression, standard
methods such as Northern blot analysis, and RT-PCR / quantitative RT-PCR are
tried-and-true. However, new advances in gene expression profiling and next-
generation sequencing can quantify gene expression levels genome-wide.
SPECIMEN COLLECTION DETAILS
(Adapted from the WHO guidelines on 2019-nCoV):

Specimen type Collection Transport Storage till testing Comment

materials to
Laboratory

Nasopharyngeal Dacron or 4 degree C < 5 days:4degreeC The nasopharyngeal

and or polyester >5 days: -70 degree and or pharyngeal
pharyngeal swab flocked C
swabs

Bronchoalveolar Sterile 4-degree c < 48 There may be some

Lavage container hours:4DegreeC dilution of pathogen
>48 hours: - but still a worthwhile
70Degree C specimen

Sputum Sterile 4-degree C <48hours:4degreeC Ensure the materials is

container >48 hours: -- from the lower
70DegreeC respiratory tract
RT-PCR
What is RT-PCR?
PCR is used widely for carrying out molecular-based diagnostic tests. It is used to
detect viruses responsible for diseases such as HIV, Ebola, African swine fever, foot-
and-mouth disease etc. RT-PCR test, and antigen tests (Serology) are two different
types of tests carried out for the diagnosis of COVID-19. The Real-Time RT–PCR is
suitable to detect the COVID-19 virus as it contains only Ribonucleic Acid. The full
form of RT-PCR is Reverse Transcriptase Polymerase Chain Reaction. It is the most
sensitive technique for mRNA detection. It quantifies the presence of a particular
genetic material of the virus or any pathogen.
Real-Time RT-PCR is loaded with fluorescent dyes as markers. Real-Time RT-PCR
helps investigators, mainly scientists or virologists, to analyse the results without
delay. RT-PCR test detects the COVID-19 virus after a nasopharyngeal swab is
collected from a patient. It detects the virus even if the viral load is less. The major
advantage of Real-Time RT-PCR is that it gives rapid results.

Procedure for RT-PCR

The following are the RT-PCR Test procedures (guidelines) in brief that are helpful
in the diagnosis of COVID-19.

Step 1: -The COVID-19 virus generally gathers in the throat or inside the nose of a
person. The RT-PCR Test diagnosis process starts with sample collection by swab
from the mentioned body parts.

Step 2: -The cells and nucleus are lysed by treating them with different chemical
solutions. Finally, the sample only contains DNA and RNA. It is a combination of
the residual genetic material of the individual and the RNA of the virus.

Step 3:- The RNA is then converted to DNA using a specific enzyme via reverse
transcription. Short fragments of DNA are also added to create a mixture.

Step 4: - If there is a virus in the sample, then the short fragments of the DNA accord
to the target divisions of the viral DNA.

Step 5:- The mixture is then put into an RT–PCR machine. The RT-PCR machine
heats and cools the mixture by cycling it through temperatures to ignite specific
chemical reactions. Through this process, the target sections of viral DNA get their
new identical copies.
Step 6: -The RT-PCR COVID-19 Test machine carries out repeated cycles with the
sample. This helps in copying the target sections of the viral DNA. At the end of each
cycle, the number of copies of the viral DNA gets doubled. As a result, about 35
billion new copies of the viral DNA sections from each strand of the virus are
generated by the end of the process.

The RT-PCR testing time is only 5-10 minutes, but the entire procedure takes up to
4 to 8 hours.

How is the result concluded?

Once the new copies of the viral DNA are formed, the marker labels then attach to

the DNA strands. The RT-PCR machine is connected to a computer that records the

amount of fluorescent dye released after each step. It displays the result on the screen

at the same time. At a certain point, when the amount of fluorescence crosses a

particular limit, this confirms the presence of the virus. A very interesting fact to

know is that the fewer the cycles to confirm the virus, the more severe is the

condition. The RT-PCR test result can be either positive or negative. The test cannot

predict when and how the virus infected you. A positive RT-PCR test means that

you are likely to be infected with COVID-19 virus.

WHY RT-PCR TEST IS BETTER THAN RAPID ANTIGEN TEST?
If you’ve gotten a coronavirus test — such as at a drive-thru or pop-up site — you
likely received a PCR test, which involved someone taking a sample from your nose
with a swab and sending it to a lab.
PCR (polymerase chain reaction tests) are highly accurate tests that work by
extracting and amplifying genetic material from the novel coronavirus,
They are much more sensitive than a rapid test, meaning PCR tests can pick up very
tiny amounts of virus so they have a high likelihood of catching an infection and not
giving a false negative.
These tests are a more expensive option, they take a more complex supply chain and
take more human labour as well as laboratory capacity to complete — all of which
are in high demand right now as everywhere.
Rapid tests like the one DeWine used are what are called antigen tests.
They can be done similar to how you would do a flu tests or a strep test you can
imagine having these in a doctor’s office or even a nursing home and to be able to
test whether someone has it in, say, 15 or 20 minutes. The disadvantage is that
they’re just not as sensitive as that molecular PCR test that we’ve been hearing
about over the last few months.
Both antigen tests and PCR tests have a place in the toolbox when it comes to
responding to the pandemic,
This method of testing is nearly immediate, which let’s contact tracing start sooner
with a virus where time is of the essence. When there’s a positive test result received
quickly, it raises the likelihood that an infectious person can be isolated before they
get a lot of other people sick.
The accuracy of available antigen tests has improved since the start of the pandemic
but they still aren’t as accurate as PCR tests. A positive antigen test for the
coronavirus is pretty trustworthy but a negative test needs treated with more
caution because the person might actually still be infectious.
That’s not something we typically see with viral PCR. That’s something that’s very
different between those two tests that has to be taken into consideration when we
look at the strategy.
The best place to use these antigen tests is large groups of population screening, he
said, where these tests can be quickly deployed and then health officials can figure
out which patients in the group need a secondary screening.
The point-of-care tests that we have available right now for COVID still suffer from
some sensitivity issues, said .And that means that when they have a negative test, if
you still have concerns for COVID or if it was still a patient that you suspected you
may have COVID, you may need to either repeat the test again or have another test
to actually confirm that it indeed was negative.
The timing of antigen tests in the disease course is important. Later in the disease,
people shed less antigen and someone with symptoms as little as seven days could
have a false negative.

PROTECTIVE WEAR AND RULES FOR LAB TECHNICIANS:

The general rules regarding Lab Safety are common for all laboratories. Also,
these rules and policies must be updated regularly. Some of the general rules
and regulations for Lab Safety in case of an emergency are provided below:

1. One must be familiar with the fire safety signs and instructions like –
Danger Fire Risk, Fire Door, Fire Exit, Fire Hose Reel, Direction Arrow,
Radiation Area, etc. and act accordingly
2. Follow the evacuation plan of the building or facility
3. Use the safety equipment as per the requirement (e.g. – the first aid kit,
fire extinguishers, wash stations, etc.)
4. For rescue operations, call the emergency phone numbers which are
displayed in the lab or building to report the situation.

5. Refer to the marking of lab equipment and areas and act accordingly. E.g.
wear protective gears before entering a radiation area or while handling
radioactive materials.
6. In case of a fire, press the alarm and go towards the fire exit sign
7. While using fire sprinklers, make sure that an area of 36-inch diameter is
kept clear.
8. Turn off all the electrical appliances in case of any emergency (e.g. – fire,
earthquake, leakage of gas, etc.)
S. No. Dress Codes Protection Gears

Wear safety glasses while working with

1. Long hair must be tied back hazardous materials

Avoid wearing jewellery and loose Make sure that you wear gloves while
2. clothes handling chemicals

Wear footwear that completely Wash your hands properly after leaving
3. covers your feet the lab

Wear face shields during the experiment

4. Avoid wearing shorts that includes heat or chemicals

SANITIZATION PROCESS IN LAb:

Cleaning and Sanitizing

Cleaning and sanitizing are two different actions, and each requires its own
specific tools and products. Cleaning removes surface soil and other
materials but usually does not remove pathogens — or not a significant enough
amount to sanitize the area. Sanitization follows cleaning and reduces or
removes the number of pathogens that are on that surface, bringing them to
levels that are safe and healthy. The entire cleaning and sanitation process
begins with cleaning, then requires rinsing, sanitization and air drying.
When it comes to laboratory sanitation, particularly when dealing with food
and beverages, the care and attention that must be given to cleaning and
sanitation are even more crucial. It is imperative that everything in the lab is
kept clean, but whenever food comes into contact with a surface, that surface
must be both cleaned and sanitized. This promotes food safety and minimizes
food borne illness as well as cross-contamination.

Waste Disposal

There are different types of waste generated in a laboratory, and each has specific
requirements for disposal. All laboratory waste is managed following a four-step
process that begins with identifying and characterizing it. Next, the waste is placed
in appropriate containers, depending on its classification, and is labeled or marked so
that it is easily identifiable. The final step is disposal that follows the protocol for its
classification. It is important that the waste is managed within its own space in the
lab. This prevents or reduces the risk of contamination of other areas or surfaces.

Laboratory waste is not only confined to contaminates or pathogens. It can be

biological waste, which is anything living such as human, animal or plant material.
If the material is infectious or harmful, it is considered to be biohazardous waste.
Glass is another type of waste that requires special handling. This can be lab
equipment like slides, beakers and other containers. A more dangerous form of
laboratory waste is sharps, or needles. Each of these poses its own threat to safety
and health.

Nonhazardous waste can be placed in a common waste container. However, other

types of waste must meet certain safety standards and placed in special containers.
Sharps are a good example of this. A sharp is anything that is able to puncture or cut
the skin. This includes scalpels, syringes, needles, razor blades and items that have a
sharp edge, like pipe that has been cut. The main concern is that the needle or glass
will puncture someone’s skin.

All materials must be contained, and waste must be disposed of. Any chemicals or
biologicals that are believed to be contaminated should be disposed of properly, in
an approved container. Typically, a leak proof, sturdy container is required. This
even means soiled cleaning and sanitizing supplies. Health and safety should always
be a priority.
STEPS FOR RT-PCR TESTING

SAMPLE COLLECTION

SPECIMEN REFFERAL FORM (SRF)

TESTING OF COLLECTED SAMPLE

AUTHORISATION

UPLOAD TO ICMR PORTAL

STEP1:
When a patient come for RT-PCR test, patient have to fill up their details in SRF forms
for rtpcr. In this form patients have to fill up their form with some basic details like Name,
Age, Sex, Phone number, Aadhar details, previous disease history etc. After fill up forms
patient hove to going to billing counter for bill & token. Then patient will go to give their
sample.

STEP 2:
After collecting samples for RT-PCR test, samples will send to the molecular biology lab
for testing along with patient history sheet. The Sample Referral form (SRF)will be done
by RT-PCR mobile app. While doing SRF there must be filled by patient details which
they are given in their history sheet like Name, Address, Sex, Age, Phone number, Aadhar
details, vaccination details, previous disease etc will be fill in the SRF mobile app for each
patients. Then a SRF id is created for each patients.

STEP 3:
When samples came to the microbiology lab for testing, Lab technicians first numbering
those sample serially then RNA extractions done by lab technician. After that RNA from
samples are send for master mix preparation room for prepare those RNA for testing in
machine. After completing the run time of the machine microbiologist, analyses graphs for
each sample and then make a decision for a positive & negative sample.
STEP 4:
After making decision of positive and negative samples by the help of graphs, patient
history sheets are going for typing and for generate a report in hospital database. After
completed typing all the patient’s history sheets are going to microbiologist for checking
the report typing and the authorised by the authorised personnel.

STEP 5:
After authorisation by microbiologist all the report including positive and negative are
uploaded to the ICMR portal.

JOB ROLE DURING TRAINING:

1. Doing SRF form entry for each patient by using RT-PCR mobile app.
2. Making serial lists of patient serial number.
3. Search patient phone number from hospital database in the case of missing
phone number
4. Upload patient RT-PCR test report in ICMR portal.

CONCLUSION:
The trainee has done his training under The Molecular biology Lab Department
in Peerless Hospitex Hospital and Research centre limited in panchasayar,
Kolkata, the trainee is thankful to the hospital authorities to give him a chance
to complete the training in this hospital. The experience acquired in this process
would be of huge help for me in future.