LAMP PCR is a nucleic acid amplification technique that uses a DNA polymerase with strand displacement activity. It requires 4-6 designed primers that recognize 6 distinct regions on the target gene. Amplification occurs isothermally without temperature cycling. It produces stem-loop DNA structures with dumbbell ends that are amplified quickly using loop primers. LAMP has advantages over PCR like higher specificity, greater detection limit, simpler operation without expensive equipment, and ability to use crude samples. It is a sensitive, specific and cost-effective technique used for diagnosing infectious diseases.

1. LAMP PCR

2. ABBREVIATION
LAMPLAMP

3. INTRODUCTION
• Solely developed by Eiken Chemical Co., Ltd.
• First reported by Notomi et al in 2000 of
EIKEN Chemical Co. Ltd., Japan. (http://www.eiken.co.jp/en/)

4. BASIC PRINCIPLE

6. Procedure of LAMP PCR
1- Design of Primers
• 4 types of primers based on the 6 distinct regions of the target
gene: the F3c, F2c and F1c regions at the 3' side and the B1, B2 and
B3 regions at the 5' side

7. Continued…
2- Amplification
• Is of two types;
A. Non-cycling Amplification
B. Cycling Amplification
A.Non-cycling Amplification:
• Generation of stem loop DNA with dumbbell-shaped structure at
both ends.
B. Cycling Amplification:
• Dumbbell-shaped DNA is quickly amplified by the use of loop
primers.

8. NON-CYCLING AMPLIFICATION
Step-1
• One of the LAMP primers anneal to the complimentary sequence
of double stranded target DNA.
• Initiates DNA synthesis using the DNA polymerase with strand
displacement activity, displacing and releasing a single stranded
DNA.
• Unlike PCR, no need for heat denaturation of ds DNA.

9. Continued…
Step-2
• Through the strand displacement
activity of DNA polymerase, a DNA
strand complementary to the
template DNA is synthesized,
starting from the 3' end of the F2
region of the FIP.
Step-3
• The F3 Primer anneals to the F3c
region, outside of FIP, on the
target DNA and
• Initiates strand displacement DNA
synthesis, releasing the FIP-linked
complementary strand.

10. Continued…
Step-4
• A double strand is formed from
the DNA strand synthesized
from the F3 Primer and the
template DNA strand.
Step-5
• The FIP-linked complementary
strand is released as a single
strand because of the
displacement by the DNA strand
synthesized from the F3 Primer.
• Released single strand forms a
stem-loop structure at the 5'
end because of the
complementary F1c and F1
regions.

11. Continued…
Step-6
• BIP-initiated DNA synthesis and
subsequent B3-primed strand
displacement DNA synthesis.
Step-7
• Double stranded DNA is produced
through the processes described in
Step-6.
Step-8
• The BIP-linked complementary strand
and forms a structure with stem-
loops dumbbell structure at each end,
serving as the starting structure for
LAMP cycling.

12. CYCLING AMPLIFICTAION

14. Continued…
3- Detection
i. Visual Detection
• Turbidity - Magnesium pyrophosphate
• Fluorescence – Calcein
ii. Gel Electrophoresis
• Lane 1 and 3 has target DNA.
• Lane 2 and 4 has non-target DNA.
• Lane M has DNA Ladder.

15. LAMP vs. PCR
• Isothermal Reaction.
• Isothermal Temperature (60-
65 C).⁰
• Doesn’t require expensive
thermocycler.
• Detection limit is greater.
• Amplification specificity is
higher as uses 4/6
oligonucleotides.
• Visualization of DNA could be
done through eyes, gel
electrophoresis and
turbidimeter.
• Cyclic Reaction.
• Variable Temperature.
Denaturation (95 C)⁰
Annealing (50-60 C)⁰
Polymerization (72 C)⁰
• Require thermocycler.
• Detection limit is lower.
• Amplification specificity is lower
than that of LAMP.
• Visualization of DNA is done
through gel electrophoresis.
Loop-mediated Isothermal
Amplification - LAMP
Polymerase Chain Reaction - PCR

16. • Could be done using crude
DNA samples.
• Loop primers accelerate
reaction rate.
• Need pure DNA samples
for amplification.
• No loop primer.
LAMP vs. PCR
Loop-mediated Isothermal
Amplification - LAMP
Polymerase Chain Reaction - PCR

17. APPLICATIONS OF LAMP

18. CONCLUSION
• LAMP is highly sensitive and specific DNA/RNA
amplification technique.
• It works on isothermal conditions and doesn't require
expensive operational machinery.
• It is simple, cost effective technique.
• LAMP is an innovation molecular diagnostic field and can
be used for the diagnosis of infectious diseases, food
inspection, environmental testing and so on.
• Innovations in biotechnology that combine molecular
biology, microfabrication and bioinformatics are moving
nucleic acid technologies from futuristic possibilities to
common laboratory techniques and modes for diagnoses.