Comparison of protein extraction methods for Mycobacterium tuberculosis detection by MALDI-TOF Mass Spectrometry
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Abstract
Tuberculosis is a respiratory disease caused by Mycobacterium tuberculosis, which is a public health problem. Currently, the MALDI-TOF mass spectrometry (MS) method has been used to detect bacteria and fungi, including M. tuberculosis. This method has high sensitivity and high specificity but there is low diagnostic cost per case when compared to other methods. However, the mycobacterial cell wall is composed of lipids that make it difficult to extract proteins from the cytoplasm. Therefore, a new protein extraction method for M. tuberculosis is developed. The ultrasound (SM) was used to increase cell wall rupture and reduce protein extraction time compared to routine methods (RM). Five samples of M. tuberculosis strain H37Ra ATCC 25177 and 32 samples of M. tuberculosis isolated from patients confirmed by real-time PCR were cultured on LJ medium for 3-4 weeks. Extraction of protein by an RM compared to using an SM. This study found that 37 protein extraction by the RM and the SM have the log score ≥ 2.000 were 72.97% and 75.67%, respectively and the log score 1.700-1.999 were 27.03% and 24.33%, respectively. The two methods are in almost perfect agreement by Kappa indexes = 1. Therefore, protein extraction of M. tuberculosis by SM can replace RM. When compared with RM, the protein extraction of M. tuberculosis by SM could reduce the sample preparation time by at least 38 minutes per sample.
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