Usefulness of Seminested Multiplex PCR in Surveillance of Imported Malaria in Spain

Rubio, José Miguel; Benito, Agustín; Berzosa, Pedro; Roche, Jesús; Puente, Sabino; Subirats, Mercedes; López‐Vélez, Rogelio; García, Luz; Alvar, Jorge

doi:10.1128/jcm.37.10.3260-3264.1999

Cited by 118 publications

(55 citation statements)

References 23 publications

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“…The detection of plasmodial DNA by molecular techniques is now well established in diagnostic situations. Polymerase chain reaction (PCR)‐based amplification techniques have been developed that have been quoted to provide relatively high levels of sensitivity in a diagnostic situation [28–30], offering a rapid and sensitive means of detection of plasmodial DNA in clinical samples. However, the use of molecular techniques to detect malarial parasites in donated blood has, for some time, been an area of debate.…”

Section: Donor and Donation Screeningmentioning

confidence: 99%

Malaria and blood transfusion

Kitchen¹,

2006

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The transmission of malaria by blood transfusion was one of the first recorded incidents of transfusion-transmitted infection. Although a number of different infections have been reported to be transmitted by transfusion since then, on a global scale malaria remains one of the most common transfusion-transmitted infections. Transfusion-transmitted malaria can have serious consequences, as infection with Plasmodium falciparum may prove rapidly fatal. Ensuring that, in non-endemic countries, the blood supply is free from malaria is problematical, especially as travel to malarious areas is increasing and there is some spread of the disease into new areas, as well as a resurgence of malaria in areas where previously it had been eradicated. In non-endemic countries, donor deferral can be effective, but clear guidelines are needed. In endemic countries the problem is far greater as the majority of donors may be potentially infected with malaria parasites. In both situations, the simple deferral of donors may be wasteful and can eventually erode the donor base. Thus, other strategies are needed to ensure safety with sufficiency. However, the screening of donations for evidence of malaria is not without its problems. Although the examination of blood films is still the basis for diagnosing acute malaria, in most situations it is not sufficiently sensitive for blood bank screening. In non-endemic countries, donor deferral in combination with screening for specific antimalarial immunoglobulin provides an effective means of minimizing the risk of transmission. In endemic countries, more specific donor questioning, consideration of seasonal variation and geographical distribution may help to identify the population of donors who are most likely to be infected. In addition, the administration of antimalarials to transfusion recipients may help to prevent transmission. Nonetheless, no matter what strategy is adopted, it is likely that cases of transfusion-transmitted malaria may still occur, so malaria must always be considered in any patient with a febrile illness post-transfusion.

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Section: Donor and Donation Screeningmentioning

confidence: 99%

Malaria and blood transfusion

Kitchen¹,

2006

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“…PCR was performed to evaluate the quality of genomic DNA extracted. A 231bp region of the small subunit of human ribosomal gene (ssrDNA) was amplified [16,17] UNR-HUF, 5’-GAGCCGCCTGGATACCGC-3’ REV, 5’-GACGGTATCTGATCGTCTTC-3’ forward and reverse primers, respectively [18].…”

Section: Methodsmentioning

confidence: 99%

Evaluation of DNA extracted from blood filter spots and eluates processed for enzyme linked immunosorbent assay (ELISA)

Xatse

Akorli

Owusu

et al. 2019

Preprint

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15Dried filter blood spots have become a significant blood collection method for screening 16 individuals for clinical purposes. When used for ELISAs, they are normally discarded after the 17 blood has been eluted. However, they may still be useful for extraction of DNA for molecular-18 based assays. The aim of this work was to determine the integrity of DNA extracted from filter 19 paper spots from which blood has initially been eluted for ELISA with sample dilution buffer 20 (SDB) and phosphate buffered saline (PBS). DNA was extracted from the eluted filter spots, the 21 eluate, and dried blood filter spots (controls) using spin column extraction. The quality and 22 quantity of the extracted DNA was assessed and used for PCR to further evaluate their 2 23 usefulness in molecular assays. Concentration of DNA obtained was dependent on the buffer 24 used for processing the filter blood blots. Accounting for the DNA concentration obtained from 25 dried blood spots, which were used as controls, DNA extracted from the already eluted blood 26 spots were 32 times higher in PBS than SDB processed filter paper. The ratio was even higher 27 for the eluates, which were 57 times higher in PBS than SDS eluates. SDB eluates had 28 significantly higher average DNA concentration than their eluted filter paper, but their purity 29 ratios were similar. 85% PCR success rate was achieved with the DNA samples. Useful DNA 30 can be extracted from blood spots after it has been eluted with SDB. Although the DNA 31 concentration and purity may be low, the DNA could be useful for rather simple PCR assays. 32 Author Summary 33Collection of blood onto filter paper has become an accepted method for screening individuals 34 for clinical and public health purposes since the 1960s. This method of blood collection has 35 become increasingly popular due to its ease and convenience in collection and transportation. 36The use of dried blood spots for clinical evaluations and research has become very significant. 37For research purposes, DBS when used for ELISAs are discarded after single use. DNA may 38 however be extracted from the used filter blots and used for molecular assays. The concentration 39 of DNA obtained may be low but simple assays like PCR could be done using the DNA 40 extracted from the eluted filter spot. 41 96 paper). DNA was extracted from the eluate, eluted filter paper and the dried unprocessed DBS 97 using the Qiagen DNA Blood and Tissue kit according to the manufacturer's instruction. DNA 98 was eluted in 150 µL of buffer AE. DNA quantity and purity were measured using Qubit ® 2.0 6 99 fluorometer (Invitrogen, Life technologies) and NanoDrop 2000c Spectrophotometer (Labtech 100 International Ltd, Thermo Scientific, UK). Samples were stored at −40°C for further analysis. 101 Amplification of human ribosomal gene 102 PCR was performed to evaluate the quality of genomic DNA extracted. A 231bp region of the 103 small subunit of human ribosomal gene (ssrDNA) was amplified [16,17] UNR-HUF, 5'-104 GAGCCGCCTGGATACCGC-3' REV, ...

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“…While conventional PCR may not be as useful for malaria diagnosis (Hänscheid and Grobusch, 2002) due to time lag in sample transportation, sample processing and result dissemination (Srinivasan et al, 2000), microfluidic PCR may prove otherwise. Microfluidic PCR can have higher sensitivity than conventional PCR (Rubio et al, 1999). Furthermore, the faster speed, adaptability to μTAS and portability of microfluidic PCR greatly boosts its usefulness in resource-scarce regions where malaria is endemic (Wang et al, 2009).…”

Section: Molecular Analysismentioning

confidence: 99%

Advances in microfluidics in combating infectious diseases

Tay

Pavesi

Yazdi

et al. 2016

Biotechnology Advances

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One of the important pursuits in science and engineering research today is to develop low-cost and user-friendly technologies to improve the health of people. Over the past decade, research efforts in microfluidics have been made to develop methods that can facilitate low-cost diagnosis of infectious diseases, especially in resource-poor settings. Here, we provide an overview of the recent advances in microfluidic devices for point-of-care (POC) diagnostics for infectious diseases and emphasis is placed on malaria, sepsis and AIDS/HIV. Other infectious diseases such as SARS, tuberculosis, and dengue are also briefly discussed. These infectious diseases are chosen as they contribute the most to disability-adjusted life-years (DALYs) lost according to the World Health Organization (WHO). The current state of research in this area is evaluated and projection toward future applications and accompanying challenges are also discussed.

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Usefulness of Seminested Multiplex PCR in Surveillance of Imported Malaria in Spain

Cited by 118 publications

References 23 publications

Malaria and blood transfusion

Malaria and blood transfusion

Evaluation of DNA extracted from blood filter spots and eluates processed for enzyme linked immunosorbent assay (ELISA)

Advances in microfluidics in combating infectious diseases

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