PCR and strain identification in Plasmodium falciparum

Wooden, Jason M.; Kyes, Sue; Sibley, Carol Hopkins

doi:10.1016/0169-4758(93)90131-x

Cited by 207 publications

(174 citation statements)

References 19 publications

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“…Preparation of DNA from filter paper was performed as previously described in detail. 22,23 Following a modified procedure used for DNA extraction from material derived from Egyptian mummies, 24 blood slides taken at days 3 or 7 after treatment were incubated in 10 mM Tris-HCl, 2 mM EDTA, 0.1% sodium dodecyl sulfate, 10 mg/ml of dithiothreitol, and 0.5 mg/ml of proteinase K for 16 hr at 37ЊC. Subsequently, the dissolved material was extracted with phenol-chloroform.…”

Section: Patients Materials and Methodsmentioning

confidence: 99%

Plasmodium falciparum: selection of serine 108 of dihydrofolate reductase during treatment of uncomplicated malaria with co-trimoxazole in Ugandan children.

Jelı́nek¹,

Kilian²,

Curtis³

et al. 1999

Am J Trop Med Hyg

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Abstract. In vivo testing for resistance of Plasmodium falciparum to co-trimoxazole (trimethoprim/sulfamethoxazole) was performed in Uganda in 41 children with uncomplicated malaria, and blood samples were screened before and after treatment for polymorphisms in the antifolate target genes for dihydrofolate reductase (DHFR) and dihydropteroate synthetase (DHPS). Selection towards a specific genotype at some codons of the DHFR and DHPS genes was observed in samples collected after exposure to co-trimoxazole drug pressure. The alleles 51-isoleucine, 59-arginine, and 108-serine of DHFR were significantly associated with clinical resistance, as was allele 581-alanine of DHPS. Resistance against antifolate combinations probably requires resistance-related polymorphisms in both the DHFR and the DHPS genes. In addition, it appears that the trimethoprim-resistant DHFR genotype differs from that for pyrimethamine at residue 108.Falciparum malaria and acute respiratory tract infection, two of the most common causes of childhood morbidity and mortality in sub-Saharan Africa, show an overlap of symptoms in a high proportion of cases. 1 The World Health Organization (WHO) recommends co-trimoxazole (trimethoprim and sulfamethoxazole) as treatment for children in malaria-endemic areas presenting with fever and respiratory symptoms. 2 This drug has been shown in the past to be effective against malaria as well as pneumonia. 3,4 The components of co-trimoxazole target selectively dihydrofolate reductase (DHFR) and dihydropteroate synthetase (DHPS), the same folate pathway enzymes as pyrimethamine/sulfadoxine, which is used in many African countries for treatment of chloroquine-resistant infections with Plasmodium falciparum. In vitro drug resistance of P. falciparum to pyrimethamine has been associated with point mutations in the gene coding for DHFR, in particular with the presence of asparagine in position 108.5 Likewise, polymorphisms at 5 highly conserved positions within DHPS have been reported in sulfadoxine-resistant isolates of P. falciparum: codons 436, 437, 581, 613, 6,7 and 540. [8][9][10] Unlike DHFR, no single polymorphism has been associated with all resistant strains and only limited data are available on the global distribution of these polymorphisms.11 In vivo testing for resistance to co-trimoxazole was performed in rural health centers in the 2 districts of Kabarole and Bundibugyo in western Uganda. Results of the clinical and parasitologic outcomes have been reported elsewhere.12 Fortyone children with symptomatic malaria from the Kabarole and Bundibugyo Districts of western Uganda were recruited for the study. Data were obtained about polymorphisms in the DHFR and DHPS genes that have been associated with antifolate resistance [5][6][7][13][14][15][16][17][18][19] by extraction and amplification of parasite DNA from filter paper and thick blood films. We report on a comparison of the prevalence of polymorphisms of the antifolate target genes at days 0 and 3/7 and their association with in vivo results in one...

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Section: Patients Materials and Methodsmentioning

confidence: 99%

Plasmodium falciparum: selection of serine 108 of dihydrofolate reductase during treatment of uncomplicated malaria with co-trimoxazole in Ugandan children.

Jelı́nek¹,

Kilian²,

Curtis³

et al. 1999

Am J Trop Med Hyg

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“…Another 50 µl were transferred to a 1.5 ml centrifuge tube in which the P. falciparum DNA was isolated using the chelex-100-iron protocol (Wooden et al 1993).…”

Section: Methodsmentioning

confidence: 99%

Evaluation of DNA Recombinant Methodologies for the Diagnosis of Plasmodium falciparum and their Comparison with the Microscopy Assay

Urdaneta¹,

Guevara²,

Jl³

1998

Mem. Inst. Oswaldo Cruz

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Section: Polymerase Chain Reaction (Pcr)mentioning

confidence: 99%

“…The PCR technique has also been used to investigate non-intestinal parasites. The effectiveness of PCR in detecting parasite species such as Leishmania and Plasmodium has already been studied (13)(14)(15)(16)(17)(18)(19)(20)(21)(22).To detect Leishmania in clinical samples, several PCR assays have been developed (17). In studies of visceral Leishmania, PCR has been used for several purposes in addition to diagnosis, such as treatment monitoring and epidemiological studies.…”

mentioning

confidence: 99%

Molecular techniques for the study and diagnosis of parasite infection

Tavares¹,

Staggemeier²,

Borges³

et al. 2011

J. Venom. Anim. Toxins incl. Trop. Dis

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Abstract:In parasitology, routine laboratory diagnosis involves conventional methods, such as optical microscopy, used for the morphological identification of parasites. Currently, molecular biology techniques are increasingly used to diagnose parasite structures in order to enhance the identification and characterization of parasites. The objective of the present study was to review the main current and new diagnostic techniques for confirmation of parasite infections, namely: polymerase chain reaction (PCR), real-time polymerase chain reaction (RT-PCR), loop-mediated isothermal amplification (LAMP), Luminex xMAP, random amplified polymorphic DNA (RAPD), amplified fragment length polymorphism (AFLP), and restriction fragment length polymorphism (RFLP), in addition to microsatellites. Molecular assays have comprehensively assisted in the diagnosis, treatment and epidemiological studies of parasitic diseases that affect people worldwide, helping to control parasitic disease mortality.Key words: parasite infection, diagnosis, molecular techniques, molecular epidemiology. Review ARticle The Journal of Venomous Animals and Toxins including Tropical Diseases ISSN 1678-9199 | 2011 | volume 17 | issue 3 | pages 239-248 INTRODUCTIONIn parasitology, routine laboratory diagnosis involves conventional methods, such as optical microscopy, used for the morphological identification of parasites (1). However, the occasional difficulty of identifying these parasite structures may decrease the sensitivity of such methods. Currently, because of these difficulties, molecular biology has been employed to detect parasites responsible for parasitic diseases (2).Traditional parasitological analyses have the advantage of being less costly without requiring expensive reagents and equipment. Additionally, such analyses can be easily performed when a trained microscopist is available. On the other hand, molecular technology demonstrates the presence of parasites based on their antigenic components or DNA segments (3). These tests are not influenced by environmental factors that usually can interfere with the results of a stool test, for example, thus ensuring highly reliable results (4).Current laboratory diagnostic methods for the identification of parasites include: polymerase chain reaction (PCR), random amplified polymorphic DNA (RAPD), amplified fragment length polymorphism (AFLP), restriction fragment length polymorphism (RFLP), microsatellite marker method, Luminex xMAP-based technology (areas of multianalyte profiling), loop-mediated isothermal amplification (LAMP), and the recently added real-time PCR (5-12).The objective of the present article was to review the applications of molecular biology techniques in parasitology by evaluating and discussing their uses and benefits. MOLECULAR METHODS FOR DETECTION OF PARASITE STRUCTURESSeveral molecular tests to detect parasites have been developed in the last decade. Their specificity and sensitivity have gradually increased, and parasites that were previously difficult to diagnose usi...

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PCR and strain identification in Plasmodium falciparum

Cited by 207 publications

References 19 publications

Plasmodium falciparum: selection of serine 108 of dihydrofolate reductase during treatment of uncomplicated malaria with co-trimoxazole in Ugandan children.

Plasmodium falciparum: selection of serine 108 of dihydrofolate reductase during treatment of uncomplicated malaria with co-trimoxazole in Ugandan children.

Evaluation of DNA Recombinant Methodologies for the Diagnosis of Plasmodium falciparum and their Comparison with the Microscopy Assay

Molecular techniques for the study and diagnosis of parasite infection

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