Isolation and Characterization of the MSP1 Genes from Plasmodium malariae and Plasmodium ovale

Birkenmeyer, Larry G.; Muerhoff, A. Scott; Dawson, George J.; Desai, Suresh M.

doi:10.4269/ajtmh.2010.09-0022

Cited by 31 publications

(51 citation statements)

References 41 publications

(49 reference statements)

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“…For P. relictum (SGS1), it was possible to identify the full 3′ end of the peptide; within this region the GPI anchor point was fully conserved at the amino acid level between P. falciparum , P. gallinaceum and P. relictum (SGS1) with the characteristic “FCSSS” motif (Figure 3). The “FCSSS” motif has been found to be extremely conserved across almost all investigated species of malariae parasites [27] and this was also the case when including more lineages of avian malariae species. In the p19 section of the avian malaria parasite peptides, it was also possible to identify the two epidermal growth factor-like domains (EGF), based on the six conserved characteristic cysteine residues [28] (Figure 3).…”

Section: Discussionmentioning

confidence: 89%

Identification and characterization of the merozoite surface protein 1 (msp1) gene in a host-generalist avian malaria parasite, Plasmodium relictum (lineages SGS1 and GRW4) with the use of blood transcriptome

Hellgren

Kutzer

Bensch

et al. 2013

Malar J

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BackgroundThe merozoite surface protein 1 (msp1) is one of the most studied vaccine candidate genes in mammalian Plasmodium spp. to have been used for investigations of epidemiology, population structures, and immunity to infections. However methodological difficulties have impeded the use of nuclear markers such as msp1 in Plasmodium parasites causing avian malaria. Data from an infection transcriptome of the host generalist avian malaria parasite Plasmodium relictum was used to identify and characterize the msp1 gene from two different isolates (mtDNA lineages SGS1 and GRW4). The aim was to investigate whether the msp1 gene in avian malaria species shares the properties of the msp1 gene in Plasmodium falciparum in terms of block variability, conserved anchor points and repeat motifs, and further to investigate the degree to which the gene might be informative in avian malaria parasites for population and epidemiological studies.MethodsReads from 454 sequencing of birds infected with avian malaria was used to develop Sanger sequencing protocols for the msp1 gene of P. relictum. Genetic variability between variable and conserved blocks of the gene was compared within and between avian malaria parasite species, including P. falciparum. Genetic variability of the msp1 gene in P. relictum was compared with six other nuclear genes and the mtDNA gene cytochrome b.ResultsThe msp1 gene of P. relictum shares the same general pattern of variable and conserved blocks as found in P. falciparum, although the variable blocks exhibited less variability than P. falciparum. The variation across the gene blocks in P. falciparum spanned from being as conserved as within species variation in P. relictum to being as variable as between the two avian malaria species (P. relictum and Plasmodium gallinaceum) in the variable blocks. In P. relictum the highly conserved p19 region of the peptide was identified, which included two epidermal growth factor-like domains and a fully conserved GPI anchor point.ConclusionThis study provides protocols for evaluation of the msp1 gene in the avian malaria generalist parasite P. relictum. The msp1 gene in avian Plasmodium shares the genetic properties seen in P. falciparum, indicating evolutionary conserved functions for the gene. The data on the variable blocks of the gene show that the msp1 gene in P. relictum might serve as a good candidate gene for future population and epidemiological studies of the parasite.

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Section: Discussionmentioning

confidence: 89%

Identification and characterization of the merozoite surface protein 1 (msp1) gene in a host-generalist avian malaria parasite, Plasmodium relictum (lineages SGS1 and GRW4) with the use of blood transcriptome

Hellgren

Kutzer

Bensch

et al. 2013

Malar J

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“…The assembled PmMSP - 1 sequences were 5160 bp (M0N00648) and 5150 bp (M0N00290 and M0N00556) in length, whereas the PoMSP - 1 sequences were both 5157 bp. Alignment of the PmMSP - 1 sequences with the full-length sequence of a Cameroonian isolate MM1A (GenBank #FJ824669) [45] revealed nucleotide and amino acid identities of ~98 and ~97%, respectively. Alignment of the PmMSP - 1 sequences from this study with the sequences of 11 Brazilian isolates by fragments showed that fragment 2 sequences corresponding to amino acids 637–858 in the MM1A sequence were the most polymorphic with 87.3–88.9 and 82.3–83.6% identities in the nucleotide and amino acid sequences, respectively (Additional file 4).…”

Section: Resultsmentioning

confidence: 99%

Plasmodium malariae and Plasmodium ovale infections in the China–Myanmar border area

Li¹,

Zhao²,

Hua

et al. 2016

Malar J

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BackgroundThe Greater Mekong Subregion is aiming to achieve regional malaria elimination by 2030. Though a shift in malaria parasite species predominance by Plasmodium vivax has been recently documented, the transmission of the two minor Plasmodium species, Plasmodium malariae and Plasmodium ovale spp., is poorly characterized in the region. This study aims to determine the prevalence of these minor species in the China–Myanmar border area and their genetic diversity.MethodsEpidemiology study was conducted during passive case detection in hospitals and clinics in Myanmar and four counties in China along the China–Myanmar border. Cross-sectional surveys were conducted in villages and camps for internally displaced persons to determine the prevalence of malaria infections. Malaria infections were diagnosed initially by microscopy and later in the laboratory using nested PCR for the SSU rRNA genes. Plasmodium malariae and P. ovale infections were confirmed by sequencing the PCR products. The P. ovale subtypes were determined by sequencing the Pocytb, Pocox1 and Pog3p genes. Parasite populations were evaluated by PCR amplification and sequencing of the MSP-1 genes. Antifolate sensitivity was assessed by sequencing the dhfr-ts and dhps genes from the P. malariae and P. ovale isolates.ResultsAnalysis of 2701 blood samples collected from the China–Myanmar border by nested PCR targeting the parasite SSU rRNA genes identified 561 malaria cases, including 161 Plasmodium falciparum, 327 P. vivax, 66 P. falciparum/P. vivax mixed infections, 4 P. malariae and 3 P. ovale spp. P. vivax and P. falciparum accounted for >60 and ~30% of all malaria cases, respectively. In comparison, the prevalence of P. malariae and P. ovale spp. was very low and only made up ~1% of all PCR-positive cases. Nevertheless, these two species were often misidentified as P. vivax infections or completely missed by microscopy even among symptomatic patients. Phylogenetic analysis of the SSU rRNA, Pocytb, Pocox1 and Pog3p genes confirmed that the three P. ovale spp. isolates belonged to the subtype P. ovale curtisi. Low-level genetic diversity was detected in the MSP-1, dhfr and dhps genes of these minor parasite species, potentially stemming from the low prevalence of these parasites preventing their mixing. Whereas most of the dhfr and dhps positions equivalent to those conferring antifolate resistance in P. falciparum and P. vivax were wild type, a new mutation S113C corresponding to the S108 position in pfdhfr was identified in two P. ovale curtisi isolates.ConclusionsThe four human malaria parasite species all occurred sympatrically at the China–Myanmar border. While P. vivax has become the predominant species, the two minor parasite species also occurred at very low prevalence but were often misidentified or missed by conventional microscopy. These minor parasite species displayed low levels of polymorphisms in the msp-1, dhfr and dhps genes.Electronic supplementary materialThe online version of this article (doi:10.1186/s12936-016-1605-y) contains ...

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“…Insertions/deletions (indels) in coding regions were determined from multiple alignments of amino acid sequences to maintain the reading frame. Sequences of the two Cameroon isolates (GenBank accession numbers FJ824670 and FJ824671) were included for comparison [14]. Tandem repeats were detected by scanning the sequence with a small window, determining the distance between exact matches and testing the statistical criteria as implemented in the Tandem Repeats Finder version 4.0 program [18].…”

Section: Methodsmentioning

confidence: 99%

“…Recently, two nucleotide sequences of the gene encoding the merozoite surface protein-1 of P. ovale curtisi ( PocMSP-1 ) from Cameroon patients were determined [14]. However, the MSP-1 sequence of P. ovale wallikeri ( PowMSP-1 ) remains unknown.…”

Section: Introductionmentioning

confidence: 99%

Low Level of Sequence Diversity at Merozoite Surface Protein-1 Locus of Plasmodium ovale curtisi and P. ovale wallikeri from Thai Isolates

2013

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BackgroundThe merozoite surface protein-1 (MSP-1) is a candidate target for the development of blood stage vaccines against malaria. Polymorphism in MSP-1 can be useful as a genetic marker for strain differentiation in malarial parasites. Although sequence diversity in the MSP-1 locus has been extensively analyzed in field isolates of Plasmodium falciparum and P. vivax, the extent of variation in its homologues in P. ovale curtisi and P. ovale wallikeri, remains unknown.Methodology/Principal FindingsAnalysis of the mitochondrial cytochrome b sequences of 10 P. ovale isolates from symptomatic malaria patients from diverse endemic areas of Thailand revealed co-existence of P. ovale curtisi (n = 5) and P. ovale wallikeri (n = 5). Direct sequencing of the PCR-amplified products encompassing the entire coding region of MSP-1 of P. ovale curtisi (PocMSP-1) and P. ovale wallikeri (PowMSP-1) has identified 3 imperfect repeated segments in the former and one in the latter. Most amino acid differences between these proteins were located in the interspecies variable domains of malarial MSP-1. Synonymous nucleotide diversity (πS) exceeded nonsynonymous nucleotide diversity (πN) for both PocMSP-1 and PowMSP-1, albeit at a non-significant level. However, when MSP-1 of both these species was considered together, πS was significantly greater than πN (p<0.0001), suggesting that purifying selection has shaped diversity at this locus prior to speciation. Phylogenetic analysis based on conserved domains has placed PocMSP-1 and PowMSP-1 in a distinct bifurcating branch that probably diverged from each other around 4.5 million years ago.Conclusion/SignificanceThe MSP-1 sequences support that P. ovale curtisi and P. ovale wallikeri are distinct species. Both species are sympatric in Thailand. The low level of sequence diversity in PocMSP-1 and PowMSP-1 among Thai isolates could stem from persistent low prevalence of these species, limiting the chance of outcrossing at this locus.

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Isolation and Characterization of the MSP1 Genes from Plasmodium malariae and Plasmodium ovale

Cited by 31 publications

References 41 publications

Identification and characterization of the merozoite surface protein 1 (msp1) gene in a host-generalist avian malaria parasite, Plasmodium relictum (lineages SGS1 and GRW4) with the use of blood transcriptome

Identification and characterization of the merozoite surface protein 1 (msp1) gene in a host-generalist avian malaria parasite, Plasmodium relictum (lineages SGS1 and GRW4) with the use of blood transcriptome

Plasmodium malariae and Plasmodium ovale infections in the China–Myanmar border area

Low Level of Sequence Diversity at Merozoite Surface Protein-1 Locus of Plasmodium ovale curtisi and P. ovale wallikeri from Thai Isolates

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