High sensitivity of detection of human malaria parasites by the use of nested polymerase chain reaction

Snounou, Georges; Viriyakosol, Suganya; Zhu, Xin Ping; Jarra, William; Pinheiro, L.; Rosário, Virgílio Estólio do; Thaithong, Sodsri; Brown, K. N.

doi:10.1016/0166-6851(93)90077-b

Cited by 1,415 publications

(1,371 citation statements)

References 8 publications

Supporting

Mentioning

1,279

Contrasting

Unclassified

Order By: Relevance

“…Although acridine orange microscopy substantially improves detection levels over Giemsa thick-film LM [11,46], PCR-based diagnostic methods targeting the small subunit rRNA (SSU rRNA) gene [47] have transformed perspectives on malaria epidemiology [48]. These assays enable detection of the four Plasmodium parasite species that infect humans at densities ~100 times lower than the limit of LM detection [47] and have evolved from simple species-specific amplification strategies to multiplex approaches that incorporate semiquantitative assessments [31,[49][50][51][52][53][54][55][56].…”

Section: Improved Diagnosis Of P Malariae and P Ovale Infections Bymentioning

confidence: 99%

“…These assays enable detection of the four Plasmodium parasite species that infect humans at densities ~100 times lower than the limit of LM detection [47] and have evolved from simple species-specific amplification strategies to multiplex approaches that incorporate semiquantitative assessments [31,[49][50][51][52][53][54][55][56]. We have greatly extended the potential use of PCR diagnosis for a wide range of epidemiological studies through our recent efforts to develop a ligase detection reaction fluorescent microsphere assay (LDR-FMA) [52,57] to evaluate all four malaria parasites of humans in a single-well multiplex format.…”

Section: Improved Diagnosis Of P Malariae and P Ovale Infections Bymentioning

confidence: 99%

See 1 more Smart Citation

Plasmodium malariae and Plasmodium ovale – the ‘bashful’ malaria parasites

Müeller

Zimmerman

Reeder

2007

Trends in Parasitology

270

245

View full text Add to dashboard Cite

Although Plasmodium malariae was first described as an infectious disease of humans by Golgi in 1886 and Plasmodium ovale identified by Stevens in 1922, there are still large gaps in our knowledge of the importance of these infections as causes of malaria in different parts of the world. They have traditionally been thought of as mild illnesses that are caused by rare and, in case of P. ovale, short-lived parasites. However, recent advances in sensitive PCR diagnosis are causing a re-evaluation of this assumption. Low-level infection seems to be common across malaria-endemic areas, often as complex mixed infections. The potential interactions of P. malariae and P. ovale with Plasmodium falciparum and Plasmodium vivax might explain some basic questions of malaria epidemiology, and understanding these interactions could have an important influence on the deployment of interventions such as malaria vaccines. Geographical distributionAlthough distribution of Plasmodium malariae infection is reported as being patchy, it has been observed in all major malaria-endemic regions of the world [1]. P. malariae infections are most common in sub-Saharan Africa and the southwest Pacific, where age-specific prevalence in mass blood surveys have exceeded 15-30% [2][3][4][5][6][7][8]. By contrast, when P. malariae has been detected in malaria-endemic regions of Asia [9][10][11][12], the Middle East [13], South America [14] and Central America [15], it is observed as an infrequent infection, with blood-smear light microscopy (LM) prevalence rarely exceeding 1-2%. Much higher levels of infection were, however, found in montagnard refugees from the Cambodian-Vietnamese border [16]. In South America, P. malariae is thought to be a zoonotic infection because the genetically identical Plasmodium brasilianum infects new-world monkeys [17] and both monkeys and humans in endemic areas show high levels of seropositivity to P. malariae and P. brasilianum antigens [18].Plasmodium ovale was thought to have a much more limited distribution, with endemic transmission traditionally described as being limited to areas of tropical Africa, New Guinea, the eastern parts of Indonesia and the Philippines [19,20]. Infections with P. ovale, however, have also been reported in the Middle East [13], the Indian subcontinent [21] and different parts of Southeast Asia [11,22,23]. In West Africa (and to a lesser extent Central Africa), age specific LM prevalence of >10% have been observed [3,6] places where P. ovale is observed, it is relatively uncommon and its prevalence (as detected by LM) rarely exceeds 3-5% [22,[24][25][26].Indepth descriptions of the epidemiology of both infections based upon LM data are, thus, restricted almost exclusively to highly endemic areas in Africa and the Southwest Pacific. Detailed epidemiological studies from South America and Asia are lacking. Variation in parasite prevalenceIn West Africa, P. malariae prevalence has been reported to peak at ages similar to those of P. falciparum (i.e. in children under ten years of...

show abstract

Section: Improved Diagnosis Of P Malariae and P Ovale Infections Bymentioning

confidence: 99%

Section: Improved Diagnosis Of P Malariae and P Ovale Infections Bymentioning

confidence: 99%

Plasmodium malariae and Plasmodium ovale – the ‘bashful’ malaria parasites

Müeller

Zimmerman

Reeder

2007

Trends in Parasitology

270

245

View full text Add to dashboard Cite

show abstract

“…However, polymorphisms resulting from point mutations have also been described in several antigens with and without regions of repeats [9]. The best studied antigens with regard to allelic polymorphisms are the merozoite surface proteins 1 and 2 (MSP-1 and MSP-2), selected regions of which are being used for genotyping of parasite populations [10].…”

Section: Introductionmentioning

confidence: 99%

Antigenic Diversity of Plasmodium falciparum and Antibody‐Mediated Parasite Neutralization

Bolad

Berzins

2000

Scand J Immunol

View full text Add to dashboard Cite

The malaria parasite Plasmodium falciparum, causing the most severe form of the disease in humans, is characterized by a broad antigenic diversity between different strains and isolates of the parasite. The antigenic diversity reflects on the one hand polymorphisms in allelic gene products and, on the other hand, antigenic variation as a result of expression of alternative genes in multigene families. Using selected polymorphic regions in two merozoite surface antigens, a method for genotyping P. falciparum parasites has been developed. This has resulted in new information on the clonal multiplicity and dynamics of parasite populations. Observations from in vivo and in vitro studies have identified many potential parasite‐neutralizing immune responses and several of the target antigens are being explored as vaccine candidates. Studies of antibody‐mediated neutralization of parasites in P. falciparum in vitro cultures, with or without leukocytes as effector cells, have been instrumental in identifying potential target antigens for protective immunity and for elucidation of the effects of immune pressure on the dynamics of parasite populations and their antigenic plasticity.

show abstract

“…Furthermore, salivary glands were considered to be free of the PCR inhibitors associated with other mosquito tissues (Schriefer et al, 1991;Snounou et al, 1993; Arez et al, 2000). We consider that these factors contributed to our success in identifying mixed species infections in wild-caught mosquitoes.…”

Section: Abstract: Salivary Glands Plasmodium Knowlesi Pcr Rt-pcrmentioning

confidence: 99%

Anopheles dirus co-infection with human and monkey malaria parasites in Vietnam

Nakazawa

Marchand²,

Quang³

et al. 2009

International Journal for Parasitology

View full text Add to dashboard Cite

The feasibility of identifying parasite DNA and specific mRNAs from wild caught Anopheles dirus mosquitoes was assessed using dried mosquito salivary glands preserved on filter paper. We were able to detect Plasmodium falciparum, Plasmodium vivax This study also shows that the preservation of mosquito salivary glands on filter paper, and the down-stream extraction of parasite DNA and RNA from those, offers a powerful resource for molecular epidemiological studies on malaria. Keywords: Salivary glands, Plasmodium knowlesi, PCR, RT-PCRMalaria transmission occurs in the forested areas in the southern and central provinces of Vietnam (Erhart et al., 2004) and constitutes a considerable public health problem in these areas. In order to fully understand the complexities of malaria transmission, it is important to identify and characterize malaria parasites in mosquito vectors. Evidence for the generation of genetic diversity during sexual proliferation in mosquito vectors has previously been accumulated using laboratory isolates of genetically diverse parasites, as well as with samples from field settings (Babiker et al., 1994; Menegon et al., 2000 knowlesi, were spotted onto chromatography-grade filter papers (ET31CHR;Whatman, Maidstone, UK). Each blood-spotted filter paper was immediately air-dried and stored in a sealed plastic bag at room temperature until RT-PCR or PCR analysis was performed. To examine the detection limit for mosquito salivary gland-extracted parasite mRNA, salivary glands from laboratory reared Anopheles stephensi were prepared in the same way as were the wild-caughtAn. dirus. Salivary glands were dissected from laboratory colony reared An.stephensi more than 1 week after blood feeding on either mouse or human blood and dried on E31CHR filter paper at room temperature. Gametocyte suspensions were obtained from cultured 3D7-9A by pyrimethamine treatment at 10 -6 M from day 7 after thawing until harvest on day 12. Ten times serially diluted cultured gametocyte suspension was added to the spot on the filter paper where the salivary glands were attached. Extraction of RNA and reverse transcription was carried out as previously described (Maeno et al., 2003). Briefly, dried spotted filter paper was cut into small pieces and total RNA and genomic DNA (gDNA) was extracted with ISOGEN (Nippon gene, Tokyo, Japan) according to the manufacturer's instructions. The extracted total RNA was transcribed to synthesize cDNA which was subsequently subjected to PCR using specific The detection limit of parasite mRNA by RT-PCR was one gametocyte per salivary gland for pfg377 mRNA and 10 gametocytes per gland for pfs16 mRNA (Fig. 1A). We extracted both parasite gDNA and mRNA from the salivary glands of wild caught mosquitoes. Region 3 of pfg377 mRNA was detected in all of the three dried salivary gland samples and both the mRNA and gDNA of pfg377 were detected in two samples (Fig. 1B). The PCR products showed the same molecular size as the RT-PCR products by electrophoresis. No PCR product was observed by ...

show abstract

High sensitivity of detection of human malaria parasites by the use of nested polymerase chain reaction

Cited by 1,415 publications

References 8 publications

Plasmodium malariae and Plasmodium ovale – the ‘bashful’ malaria parasites

Plasmodium malariae and Plasmodium ovale – the ‘bashful’ malaria parasites

Antigenic Diversity of Plasmodium falciparum and Antibody‐Mediated Parasite Neutralization

Anopheles dirus co-infection with human and monkey malaria parasites in Vietnam

Contact Info

Product

Resources

About