Background
Plasmodium falciparum
merozoite surface protein-1 (PfMSP-1) and -2 (PfMSP-2) are major blood-stage vaccine candidate antigens. Understanding the genetic diversity of the genes,
pfmsp
-
1
and
pfmsp
-
2
, is important for recognizing the genetic structure of
P. falciparum
, and the development of an effective vaccine based on the antigens. In this study, the genetic diversities of
pfmsp
-
1
and
pfmsp
-
2
in the Myanmar
P. falciparum
were analysed.
Methods
The
pfmsp
-
1
block 2 and
pfmsp
-
2
block 3 regions were amplified by polymerase chain reaction from blood samples collected from Myanmar patients who were infected with
P. falciparum
in 2013–2015. The amplified gene fragments were cloned into a T&A vector, and sequenced. Sequence analysis of Myanmar
pfmsp
-
1
block 2 and
pfmsp
-
2
block 3 was performed to identify the genetic diversity of the regions. The temporal genetic changes of both
pfmsp
-
1
and
pfmsp
-
2
in the Myanmar
P. falciparum
population, as well as the polymorphic diversity in the publicly available global
pfmsp
-
1
and
pfmsp
-
2
, were also comparatively analysed.
Results
High levels of genetic diversity of
pfmsp
-
1
and
pfmsp
-
2
were observed in the Myanmar
P. falciparum
isolates. Twenty-eight different alleles of
pfmsp
-
1
(8 for K1 type, 14 for MAD20 type, and 6 for RO33 type) and 59 distinct alleles of
pfmsp
-
2
(18 for FC27, and 41 for 3D7 type) were identified in the Myanmar
P. falciparum
population in amino acid level. Comparative analyses of the genetic diversity of the Myanmar
pfmsp
-
1
and
pfmsp
-
2
alleles in the recent (2013–2015) and past (2004–2006) Myanmar
P. falciparum
populations indicated the dynamic genetic expansion of the
pfmsp
-
1
and
pfmsp
-
...
The Beijing genotype of Mycobacterium tuberculosis is known to be a worldwide epidemic clade. It is suggested to be a possibly resistant clone against BCG vaccination and is also suggested to be highly pathogenic and prone to becoming drug resistant. Thus, monitoring the prevalence of this lineage seems to be important for the proper control of tuberculosis. The Rv0679c protein of M. tuberculosis has been predicted to be one of the outer membrane proteins and is suggested to contribute to host cell invasion. Here, we conducted a sequence analysis of the Rv0679c gene using clinical isolates and found that a single nucleotide polymorphism, C to G at position 426, can be observed only in the isolates that are identified as members of the Beijing genotype family. Here, we developed a simple multiplex PCR assay to detect this point mutation and applied it to 619 clinical isolates. The method successfully distinguished Beijing lineage clones from non-Beijing strains with 100% accuracy. This simple, quick, and cost-effective multiplex PCR assay can be used for a survey or for monitoring the prevalence of Beijing genotype M. tuberculosis strains.
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