LAMP, simple, fast, and accurate alternative method of TB diagnosis

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August 10, 2020
September 5, 2020

LAMP, simple, fast, and accurate alternative method of TB diagnosis

Tuberculosis (TBC) is still a serious global health problem. It is estimated that 10 million people are infected with TB every year worldwide. More than 95% of deaths from tuberculosis occur in developing countries. Indonesia is one of the countries with the most TB problems (high burden country)) and was ranked third after India and China. It is estimated that there were 845,000 TB cases in Indonesia in 2018 and 24,000 of them were drug resistant TB cases. The main causes of the high TB cases are poverty, less optimal TB treatment programs and demographic changes due to an increasing population. Besides its treatment, diagnosis of TB has an important role in overcoming this problem. In a simple method, the diagnosis of TB can be done by looking at the sputum smear using a microscope. However, this method has many obstacles in its implementation such as the lack of trained personnel and the subjectivity of the final result which is very much determined by the accuracy and expertise of the laboratory staff. Therefore, another method which is easier to work with and is faster, more sensitive is needed so TB cases can be reduced.

The loop-mediated isothermal amplification (LAMP) is one method that can be used for the diagnosis of tuberculosis. LAMP uses the isothermal principle, at a temperature of 64 o C, to carry out the DNA copying reaction without the need for a thermal cycler machine such as the polymerization method (PCR), LAMP only requires a water bath or heat block that is simpler so that it can be used as a standard diagnosis as it is easy to work with in health service units including Puskesmas (community healthcare center). Although bacterial culture is the standard recommended by WHO for diagnosis of TB, this method is slow and expensive, making it difficult to become a widespread diagnostic tool. Meanwhile, LAMP is fast diagnostic method with the advantages of accuracy on par with bacterial culture method.

Which one is better between the LAMP method or the microscope use in the diagnosis of TB? The results of this study indicate that the LAMP method has better sensitivity than the microscopic method. It is also supported by the results of previous studies. The LAMP method only requires a temperature of 64 oC for one hour to carry out the DNA copying reaction effectively and efficiently. Visualization of results can also be done with the naked eye because the color change can be directly compared between positive and negative results. Determination of results using microscopic methods requires special skills in observing the number of bacteria in the prepared slides, the smear that is made also determines the result of the diagnosis, not all bacteria contained in the sputum sample can be caught in the 2 cm x 3 cm slides commonly used in microscopic method, so the final result tends to be subjective.

A precise, fast, and accurate diagnosis is the first door to success in the treatment of diseases caused by Mycobacterium tuberculosis species, which are still claiming many victims. LAMP can be a new alternative in the TB diagnosis process to support end TB programs launched by WHO, especially in developing countries with technological limitations. LAMP method is simple without the need of a special machine used as a reference in confirming the TB diagnosis results with the same quality as the PCR method. Due to its simplicity and effectiveness, the LAMP method also allows it to be used in emergencies in the field as long as it maintains occupational safety and security standards.

Author: Ni Njoman Juliasih (ITD UNAIR)

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