scholarly journals Sequence Analysis of the K13-Propeller Gene in Artemisinin Challenging Plasmodium falciparum Isolates from Malaria Endemic Areas of Odisha, India: A Molecular Surveillance Study

2020 ◽  
Vol 2020 ◽  
pp. 1-6
Author(s):  
Ramakanta Rana ◽  
Manoranjan Ranjit ◽  
Madhusmita Bal ◽  
Hemant Kumar Khuntia ◽  
Sanghamitra Pati ◽  
...  
Keyword(s):  
Plasmodium Falciparum ◽  
Hot Spots ◽  
Reference Sequence ◽  
Nested Polymerase Chain Reaction ◽  
Molecular Surveillance ◽  
Base Pair Fragment ◽  
Baseline Information ◽  
Polymerase Chain ◽  
Dna Alignment ◽  
Pair Fragment

Estimation of the spread and advancement of Plasmodium falciparum artemisinin-resistant parasites can be done by probing polymorphisms in the kelch (Pfk13) domain (a validated molecular marker). This study aimed to provide baseline information for future artemisinin surveillance by analyzing the k13-propeller domain in P. falciparum field isolates collected from 24 study areas in 14 malaria hot spots of Odisha (previously Orissa) during July 2018-January 2019. A total of 178 P. falciparum mono infections were assessed. An 849-base pair fragment encoding the Pfk13 propeller was amplified by nested polymerase chain reaction and sequenced in both directions (PCR). After DNA alignment with the 3D7 reference sequence, all samples were found to be wild type. It can be anticipated that malaria public health is not under direct threat in Odisha relating to ART resistance.

Detection of Plasmodium falciparum in Human Blood by a Nested Polymerase Chain Reaction

1994 ◽  
Vol 51 (5) ◽  
pp. 617-626 ◽  
Author(s):  
Meiji Arai ◽  
Yusuke Wataya ◽  
Toshifumi Kakutani ◽  
Chiyoko Mizukoshi ◽  
Fumie Kubochi
Keyword(s):  
Polymerase Chain Reaction ◽  
Plasmodium Falciparum ◽  
Human Blood ◽  
Chain Reaction ◽  
Nested Polymerase Chain Reaction ◽  
Polymerase Chain

scholarly journals Identification of Sugarcane Striate Mosaic-Associated Virus by Partial Characterization of Its Double-Stranded RNA

1999 ◽  
Vol 89 (10) ◽  
pp. 877-883 ◽  
Author(s):  
Yoon Gi Choi ◽  
Barry J. Croft ◽  
John W. Randles
Keyword(s):  
Double Stranded Rna ◽  
Virus Particles ◽  
Reverse Transcription Pcr ◽  
Protein Size ◽  
Putative Coat Protein ◽  
Base Pair Fragment ◽  
Potato Virus M ◽  
Polymerase Chain ◽  
Pair Fragment

Sugarcane striate mosaic (ScSM)-affected sugarcane leaves contain a disease-associated 9-kilobase (kb) double-stranded RNA (dsRNA), usually together with 6- and 2.6-kb dsRNAs. The purified 9-kb dsRNA was amplified by the randomly primed polymerase chain reaction (PCR) and cloned. The nucleotide sequences of three separate regions, representing about 2.55 kb (28%) of the dsRNA sequence, were found to have significant similarities to viruses in the genera Capillo-, Carla-, Fovea-, Potex-, Poty-, Tricho-, and Tymovirus. Greatest overall similarity was found to apple stem pitting virus, with less similarity to blueberry scorch virus and potato virus M. A standard virus purification procedure was used to identify slightly flexuous filamentous particles that copurified with the disease-associated RNA. Particle modal lengths were approximately 950 and 1,900 nm with a diameter of 15 nm. Preparations contained a 51-kDa putative capsid protein and a 9-kb single-stranded RNA with a probable 3′ polyadenylate tract. These ScSM-associated virus particles differ physically from viruses in existing genera because of their relative rigidity, length, and putative coat protein size. Reverse-transcription PCR with a primer pair designed from the sequenced segments amplified a 820-base pair fragment from ScSM-affected but not healthy sugarcane plants.

Detection of Theileria annulata in cattle and vector ticks by PCR using the Tams1 gene sequences

Parasitology ◽  
2000 ◽  
Vol 120 (3) ◽  
pp. 245-254 ◽  
Author(s):  
E. KIRVAR ◽  
T. ILHAN ◽  
F. KATZER ◽  
P. HOOSHMAND-RAD ◽  
E. ZWEYGARTH ◽  
...  
Keyword(s):  
Blot Hybridization ◽  
Southern Blot Hybridization ◽  
Theileria Annulata ◽  
Babesia Bigemina ◽  
Chain Reaction ◽  
Hyalomma Anatolicum Anatolicum ◽  
Merozoite Surface Antigen ◽  
Base Pair Fragment ◽  
Polymerase Chain ◽  
Pair Fragment

A Polymerase Chain Reaction (PCR) and Southern blot hybridization for the detection of Theileria annulata are described. The PCR used primers amplifying a 785 base-pair fragment of the T. annulata gene which encodes the 30 kDa major merozoite surface antigen, Tams1. The sensitivity of the PCR in bovine blood was 1 piroplasm in 1 μl of blood. T. buffeli, T. parva, Babesia bigemina, B. bovis and B. divergens were not detected. The PCR detected down to 1 infected acinus/tick in resting and partially fed adult Hyalomma anatolicum anatolicum ticks and was negative for T. lestoquardi and T. equi, which are transmitted by this tick but are not infective to cattle. The specificity of the PCR was checked using 30 stocks of T. annulata, all of which were detected. Three stocks of T. lestoquardi, 4 of T. equi and 1 each of T. buffeli, T. parva, B. bigemina, B. bovis and B. divergens were used to ascertain there were no cross-reactions. A nested PCR using separate primers for the first reaction and the same primers for the second reaction detected T. annulata to the same sensitivity and specificity in saponin-extracted DNA samples stored for long periods at −20 °C.

scholarly journals Genetic variations in histidine-rich protein 2 and histidine-rich protein 3 of Myanmar Plasmodium falciparum isolates

2020 ◽  
Vol 19 (1) ◽  
Author(s):  
Hương Giang Lê ◽  
Jung-Mi Kang ◽  
Jinyoung Lee ◽  
Won Gi Yoo ◽  
Moe Kyaw Myint ◽  
...  
Keyword(s):  
Plasmodium Falciparum ◽  
Genetic Variation ◽  
Comparative Analysis ◽  
Amino Acid ◽  
Structural Features ◽  
Nested Polymerase Chain Reaction ◽  
Blood Samples ◽  
Malaria Rapid Diagnostic Tests ◽  
Length Polymorphisms ◽  
Polymerase Chain

Abstract Background Malaria rapid diagnostic tests (RDTs) are precious tools to diagnose malaria. Most RDTs used currently are based on the detection of Plasmodium falciparum histidine-rich protein 2 (PfHRP2) in a patient’s blood. However, concern has been raised in recent years that deletion of pfhrp2 in the parasite could affect the accuracy of PfHRP2-based RDTs. In addition, genetic variation in pfhrp2 might influence the accuracy and sensitivity of RDTs. In this study, the genetic variation in pfhrp2 and pfhrp3 in Myanmar P. falciparum isolates was analysed. Methods Blood samples were collected from malaria patients who were infected with P. falciparum in Mandalay, Naung Cho, Tha Beik Kyin, and Pyin Oo Lwin, Upper Myanmar between 2013 and 2015. The pfhrp2 and pfhrp3 were amplified by nested polymerase chain reaction (PCR), cloned and sequenced. Genetic variation in Myanmar pfhrp2 and pfhrp3 was analysed using the DNASTAR program. Comparative analysis of Myanmar and global pfhrp2 and pfhrp3 isolates was also performed. Results One-hundred and two pfhrp2 and 89 pfhrp3 were amplified from 105 blood samples, of which 84 pfhrp2 and 56 pfhrp3 sequences were obtained successfully. Myanmar pfhrp2 and pfhrp3 showed high levels of genetic variation with different arrangements of distinct repeat types, which further classified Myanmar pfhrp2 and pfhrp3 into 76 and 47 haplotypes, respectively. Novel amino acid changes were also found in Myanmar pfhrp2 and pfhrp3, but their frequencies were very low. Similar structural organization was shared by Myanmar and global pfhrp2 and pfhrp3, and differences in frequencies of repeat types and lengths were also observed between and among global isolates. Conclusion Length polymorphisms and amino acid substitutions generated extensive genetic variation in Myanmar pfhrp2 and pfhrp3. Comparative analysis revealed that global pfhrp2 and pfhrp3 share similar structural features, as well as extensive length polymorphisms and distinct organizations of repeat types. These results provide a better understanding of the genetic structure of pfhrp2 and pfhrp3 in global P. falciparum populations and suggest useful information to develop RDTs with improved quality.

Rapid and sensitive identification ofNeospora caninumbyin vitroamplification of the internal transcribed spacer 1

Parasitology ◽  
1996 ◽  
Vol 112 (2) ◽  
pp. 177-182 ◽  
Author(s):  
O. J. M. Holmdahl ◽  
J. G. Mattsson
Keyword(s):  
Internal Transcribed Spacer ◽  
Neospora Caninum ◽  
Neuromuscular Disorders ◽  
Rrna Gene ◽  
Tissue Samples ◽  
Rdna Unit ◽  
Coccidian Parasites ◽  
Base Pair Fragment ◽  
Polymerase Chain ◽  
Pair Fragment

SummaryNeospora caninumandN. caninum-like organisms are cyst-forming coccidian parasites known to cause neuromuscular disorders in dogs and abortion in cattle. In this article we report on the use of the polymerase chain reaction (PCR) for the detection of DNA fromN. caninum. After determining the sequence of the internal transcribed spacer 1 (ITSl) ofN. caninumandToxoplasma gondii, and part of the sequences for & species ofSarcocystis, we designed a primer set for the amplification of a 279-base-pair fragment of ITSl fromN. caninum. The PCR system made possible the specific detection of 5N. caninumorganisms and no amplification was observed from any of the other cyst-forming coccidia tested, including the closely relatedT. gondii. Furthermore, we were also able to demonstrate the presence ofN. caninumin brain and lung tissue samples from experimentally infected mice. Our data also link the 5-8S rRNA gene forT. gondiiandN. caninumto the 16S-like rRNA gene, within the rDNA unit.

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