PCR-RFLP for Rapid Subtyping of Plasmodium vivax Korean Isolates

AbstractPlasmodium vivax is the most common cause of malaria worldwide as well as southeastern Turkey. After the implementation of a successful national elimination program that the local malaria cases were not reported in 2011, malaria returned to county of Savur located in southeastern Turkey in summer of 2012. The present study aimed to determine the prevalent P. vivax genotypes isolated from southeastern Turkey. Genetic polymorphism in P. vivax CSP gene was analyzed by PCR-RFLP to assess the ratio of VK210 and VK247 types. Blood samples were obtained from 15 patients who lived in southeastern between 2005-2006. According to the results, VK210 type was detected in 10 samples (66.6%), VK247 type was observed in three samples (20%). Remaining two samples showed mixed infection (13.3%). The results of the present study first time showed the ratio of P. vivax genotypes in southeastern Turkey before the elimination in 2011. The results of the present study will be enable researchers to compare the new isolates with the previously detected ones and design new treatment and/elimination strategies.

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PCR/RFLP-Based Analysis of Genetically Distinct Plasmodium vivax Population of Pvmsp-3α and Pvmsp-3β genes in Pakistan

Malaria Journal ◽

10.1186/1475-2875-13-355 ◽

2014 ◽

Vol 13 (1) ◽

Cited By ~ 4

Author(s):

Shahid Niaz Khan ◽

Asif Khan ◽

Sanaullah Khan ◽

Sultan Ayaz ◽

Sobia Attaullah ◽

...

Keyword(s):

Plasmodium Vivax ◽

Vivax Population ◽

Pcr Rflp

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Antirelapse Efficacy of Various Primaquine Regimens for Plasmodium vivax

Malaria Research and Treatment ◽

10.1155/2014/347018 ◽

2014 ◽

Vol 2014 ◽

pp. 1-11 ◽

Cited By ~ 4

Author(s):

D. D. Rajgor ◽

N. J. Gogtay ◽

V. S. Kadam ◽

M. M. Kocharekar ◽

M. S. Parulekar ◽

...

Keyword(s):

Plasmodium Vivax ◽

Recurrence Rate ◽

Relapse Rate ◽

Continuous Monitoring ◽

Standard Dose ◽

Open Label ◽

Randomized Controlled ◽

Parallel Group ◽

Pcr Rflp ◽

Group A

Background. Efficacy of standard dose of primaquine (PQ) as antirelapse for P. vivax has decreased. We aimed to assess efficacy of different PQ regimens. Methods. It was an open label, randomized, controlled, parallel group, assessor blind study comparing antirelapse efficacy of 3 PQ regimens (B=15 mg/day × 14 days, C=30 mg/day × 7 days, and D=30 mg/day × 14 days) with no PQ group (A) in P. vivax patients. Paired primary and recurrence samples were subjected to 3 methods: (i) month of recurrence and genotyping, (ii) by PCR-RFLP, and (iii) PCR sequencing, to differentiate relapse and reinfection. The rates of recurrence relapse and reinfection were compared. Methods were compared for concordance between them. Results. The recurrence rate was 16.39%, 8.07%, 10.07%, and 6.62% in groups A, B, C, and D, respectively P=0.004. The relapse rate was 6.89%, 1.55%, 4%, and 3.85% as per the month of recurrence; 8.2%, 2%, 4.58%, and 3.68% P=0.007 as per PCR-RFLP; and 2.73%, 1.47%, 1.55%, and 1.53% as per PCR sequencing for groups A, B, C, and D, respectively. The concordance between methods was low, 45%. Conclusion. The higher recurrence rate in no PQ as compared to PQ groups documents PQ antirelapse activity. Regimens tested were safe. However, probable resistance to PQ warrants continuous monitoring and low concordance and limitations in the methods warrant caution in interpreting.

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Genotipificación del gen MSP3 en pacientes con Malaria complicada

Revista avances en salud ◽

10.21897/25394622.1391 ◽

2017 ◽

pp. 16-25

Author(s):

Yeiner Espitia D ◽

Carlos Castro C ◽

Virginia Rodríguez R ◽

Luis Causil ◽

María Camila Velasco P ◽

...

Keyword(s):

Plasmodium Vivax ◽

Chelex 100 ◽

Pcr Rflp

Objetivo. Se estudió la diversidad genética de las poblaciones naturales de parásitos de Plasmodium vivax mediante el estudio de la región variable del gen Pvmsp-3α. Materiales y métodos. Se realizó PCR-RFLP en 29 muestras de pacientes con malaria por P. vivax con complicaciones (PC) y 32 pacientes con malaria por P. vivax sin complicaciones (PNC), todos los pacientes provenientes de la región Bajo-Córdoba-Uraba, zona altamente endémica para malaria. Fueron colectadas muestras de sangre total para la extracción de DNA utilizando la tecnica saponina/chelex-100. Se usó la tecnica de RFLP usando las enzimas de restricción HhAI y AluI. Diez patrones de restricción fácilmente distinguibles, se detectaron después que los productos de la PCR fueron digeridos con la enzima Alu I y 9 con la enzima Hha I. Resultados. El gen Pvmsp-3α exhibió gran polimorfismo y los resultados sugieren que este gen puede ser utilizado en Colombia como un marcador molecular epidemiológico para el genotipado de P. vivax, ademas el patron PH1con la enzima AluI mostró mayor frecencia en los pacientes complicados. Conclusiones. El gen Pvmsp3α puede ser un marcador genetico de variabilidad en las cepas circulantes de P. vivax y el PH1 sugiere ser un marcador para predecir una complicación en pacientes con malaria vivax.

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Polymorphism in the IL-1β promoter is associated with IgG antibody response to circumsporozoite protein repeats of Plasmodium vivax

Transactions of the Royal Society of Tropical Medicine and Hygiene ◽

10.1093/trstmh/traa055 ◽

2020 ◽

Vol 114 (11) ◽

pp. 858-865

Author(s):

Marcela Petrolini Capobianco ◽

Gustavo Capatti Cassiano ◽

Luciane Moreno Storti-Melo ◽

Tamirys Simão Pimenta ◽

Ana Paula Drummond Rodrigues ◽

...

Keyword(s):

Antibody Response ◽

Plasmodium Vivax ◽

Circumsporozoite Protein ◽

Igg Antibody ◽

Specific Pcr ◽

Pcr Rflp ◽

Allele Specific ◽

Study Association ◽

Antibody Levels ◽

Host Parasite

Abstract Background It is well established that infection by Plasmodium vivax is a result of host-parasite interactions. In the present study, association with the IL1/IL2 cytokine profiles, anticircumsporozoite protein antibody levels and parasitic loads was evaluated in individuals naturally infected with P. vivax in an endemic area of the Brazilian Amazon. Methods Molecular diagnosis of P. vivax and variants was performed using the PCR-RFLP method and IL1B -511C>T, IL2 -330T>G and IL2+114T>G polymorphisms were identified using PCR-RFLP and allele-specific PCR. IL-1β and IL-2 cytokine levels were detected by flow cytometry and circumsporozoite protein (CSP) antibodies were measured by ELISA. Results Three variants of P. vivax CSP were identified and VK247 was found to be the most frequent. However, the prevalence and magnitude of IgG antibodies were higher for the VK210 variant. Furthermore, the antibody response to the CSP variants was not associated with the presence of the variant in the infection. Significant differences were observed between the single nucleotide polymorphism (SNP) -511T>C in the IL1B gene and levels of antibodies to the VK247 and P. vivax-like variants, but there were no associations between SNPs in IL1 and IL2 genes and their plasma products. Conclusions Individuals with the rs16944 CC genotype in the IL1β gene have higher antibody levels to the CSP of P. vivax of VK247 and P. vivax-like variants.

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PCR-RFLP Based genetic diversity of Plasmodium vivax genotypes in district Mardan, Pakistan

Brazilian Journal of Biology ◽

10.1590/1519-6984.241110 ◽

2022 ◽

Vol 82 ◽

Author(s):

I. Ullah ◽

S. G. Afridi ◽

A. U. Khan ◽

M. Israr ◽

A. Ali ◽

...

Keyword(s):

Genetic Diversity ◽

Plasmodium Vivax ◽

Nested Pcr ◽

Vaccine Development ◽

Restriction Enzymes ◽

Type A ◽

P Value ◽

Type B ◽

Pcr Rflp ◽

Type C

Abstract Plasmodium vivax is the most common human malaria parasite in Asian countries including Pakistan. Present study was designed to explore the genetic diversity of plasmodium vivax genotypes based on Pvmsp-3α and Pvmsp-3βgenes using allelic specific nested PCR and RFLP assays markers from field isolates in district Mardan, Pakistan. Blood samples of 200 P. vivax malarial patients were collected after taking their written informed consent. Genetic diversity in nested PCR products was determined by Restriction Fragment Length Polymorphism (RFLP) utilizing Alu1 and PstI restriction enzymes for alpha and beta gene products digestion, respectively. For analysis the genetic diversity of the sub allelic variants of Pvmsp3α and Pvmsp3β genes, Chi-Square test was performed by utilizing Minitab programming software 18. The P value 0.05 was considered as statistically significant. For Pvmsp-3α genes after gel electrophoresis of digested products, four distinct genotypes were obtained from total of 50 samples; type A: 35 (70%) (1.5-2.0 kb), 12 of type B (24%) (1.5-1.7 kb), 2 of type C (4%) (0.5-1.5) and one for type D (2%) (0.5-0.65 kb) which could be characterized into 9 allelic pattern (A1-A4, B1-B3, C1, D), in which A3 remained the most predominant. For Pvmsp-3βgenes, three distinct genotypes were obtained from 50 samples; 40(80%) of type A (1.5-2.5 kb), 9 (18%) of type B (1.0-1.5kb) and 1(2%) of type C (0.65 kb) which could be characterized into 6 allelic patterns (A1-A3, B1-B2, and C1). Most dominant one in Type A was A1 alleles which were noted (46%), while in Type B, the most dominant were B1 (10%).This study is the first ever report of molecular epidemiology and genetic variation in Pvmsp-3α and Pvmsp-3β genes of P. vivax isolates by using PCR/RFLP from District Mardan and showed a remarkable level of genetic diversity in the studied genes of circulating parasites in the study area. The results of this study will contribute in future studies about the genetic structure of parasite and vaccine development against the malaria.

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Detection of Plasmodium vivax in a liver sample of a howler-monkey: one evidence more in favour of the identity between Plasmodium simium and P. vivax

10.1101/2020.09.06.279869 ◽

2020 ◽

Author(s):

J.C. Buery ◽

A.M.R.C. Duarte ◽

F.E.C. Alencar ◽

C. Furieri ◽

S.L. Mendes ◽

...

Keyword(s):

Plasmodium Vivax ◽

Atlantic Forest ◽

Low Frequency ◽

Plasmodium Malariae ◽

Spatial Distance ◽

Howler Monkey ◽

Liver Sample ◽

Zoonotic Malaria ◽

Zoonotic Parasite ◽

Pcr Rflp

ABSTRACTIntroductionThe residual malaria of Atlantic Forest systems in Brazil occurs as an endemic disease with low frequency of cases. The chronological and spatial distance among the cases indicate an absence of fitness to the classical malaria cycle. This peculiar condition raised the suspicion of a reservoir, possibly the non-human primates. Simian and human malaria occur at the same places in that region, and there is already evidence of molecular identity between the simian parasites, Plasmodium simium and Plasmodium brasilianum, and the human parasites, Plasmodium vivax and Plasmodium malariae, respectively. Two different SNPs identified in the COX1 region of the Plasmodium vivax/simium of the Atlantic Forest reinforced its characterization as a zoonotic parasite. This finding supported the development of a PCR-RFLP protocol to identify such polymorphisms, and to monitor zoonotic malaria transmission.MethodsIn the present work, we tested the above-mentioned PCR-RFLP protocol in unprecedented mosquitoes and simian samples collected in Espírito Santo State, Brazil (ES).ResultsThe parasite found in the simian sample was P. vivax, contrary to what the protocol should indicate. In the mosquito samples, the protocol disclosed both forms of the parasite.ConclusionThis result suggests that the previously published pair of SNPs, and, consequently, the PCR-RFLP protocol, are not able to distinguish the dynamics of Plasmodium spp. circulation in the Atlantic Forest endemic area of ES.

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Genetic structures of geographically distinct Plasmodium vivax populations assessed by PCR/RFLP analysis of the merozoite surface protein 3β gene

Acta Tropica ◽

10.1016/j.actatropica.2006.10.011 ◽

2006 ◽

Vol 100 (3) ◽

pp. 205-212 ◽

Cited By ~ 24

Author(s):

Zhaoqing Yang ◽

Jun Miao ◽

Yaming Huang ◽

Xinyi Li ◽

Chaturong Putaporntip ◽

...

Keyword(s):

Plasmodium Vivax ◽

Surface Protein ◽

Rflp Analysis ◽

Merozoite Surface Protein ◽

Pcr Rflp ◽

Genetic Structures

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Molecular authentication of Thai medicinal plant Khamin khruea by PCR-RFLP analysis

Planta Medica ◽

10.1055/s-0028-1084538 ◽

2008 ◽

Vol 74 (09) ◽

Author(s):

P Rojsanga ◽

W Gritsanapan ◽

W Leelamanit ◽

S Sukrong

Keyword(s):

Medicinal Plant ◽

Rflp Analysis ◽

Molecular Authentication ◽

Pcr Rflp

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Simple and Rapid Detection of Factor V Leiden by Allele-specific PCR Amplification

Thrombosis and Haemostasis ◽

10.1055/s-0038-1650362 ◽

1996 ◽

Vol 75 (05) ◽

pp. 757-759 ◽

Cited By ~ 11

Author(s):

Rainer Blasczyk ◽

Markus Ritter ◽

Christian Thiede ◽

Jenny Wehling ◽

Günter Hintz ◽

...

Keyword(s):

Direct Sequencing ◽

Activated Protein C ◽

Factor V Leiden ◽

Pcr Amplification ◽

Factor V ◽

Specific Pcr ◽

Pcr Rflp ◽

Allele Specific ◽

Specific Amplification ◽

Allele Specific Pcr

SummaryResistance to activated protein C is the most common hereditary cause for thrombosis and significantly linked to factor V Leiden. In this study, primers were designed to identify the factor V mutation by allele-specific PCR amplification. 126 patients with thromboembolic events were analysed using this technique, PCR-RFLP and direct sequencing. The concordance between these techniques was 100%. In 27 patients a heterozygous factor VGln506 mutation was detected, whereas one patient with recurrent thromboembolism was homozygous for the point mutation. Due to its time- and cost-saving features allele-specific amplification should be considered for screening of factor VGln506.

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