Correction for Cutts et al., “Inactivating Zaire Ebolavirus in Whole-Blood Thin Smears Used for Malaria Diagnosis”

Malaria is an important mimic or coinfection in potential Ebolavirus disease patients. Here, we evaluated the efficacy of the 100% methanol-inactivating Zaire Ebolavirus Makona variant for malaria thin-smear preparation. We determined that 100% methanol completely inactivated the virus after 15 min.

Download Full-text

An Alternative Method for Extracting Plasmodium DNA from EDTA Whole Blood for Malaria Diagnosis

The Korean Journal of Parasitology ◽

10.3347/kjp.2018.56.1.25 ◽

2018 ◽

Vol 56 (1) ◽

pp. 25-32 ◽

Cited By ~ 1

Author(s):

Krongkaew Seesui ◽

Kanokwan Imtawil ◽

Phimphakon Chanetmahun ◽

Porntip Laummaunwai ◽

Thidarut Boonmars

Keyword(s):

Whole Blood ◽

Malaria Diagnosis ◽

Alternative Method

Download Full-text

Comparative performance of PCR using DNA extracted from dried blood spots and whole blood samples for malaria diagnosis: a meta-analysis

Scientific Reports ◽

10.1038/s41598-021-83977-5 ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Aongart Mahittikorn ◽

Frederick Ramirez Masangkay ◽

Kwuntida Uthaisar Kotepui ◽

Giovanni De Jesus Milanez ◽

Manas Kotepui

Keyword(s):

Whole Blood ◽

Large Scale ◽

Meta Analysis ◽

Malaria Diagnosis ◽

Analysis Data ◽

Dried Blood Spots ◽

Epidemiological Studies ◽

Comparative Performance ◽

Blood Spots ◽

Significant Difference

AbstractPolymerase chain reaction (PCR) using deoxyribonucleic acid (DNA) extracted from dried blood spots (DBS) provides a fast, inexpensive, and convenient method for large-scale epidemiological studies. This study compared the performance of PCR between DNA extracted from DBS and DNA obtained from whole blood for detecting malarial parasites. Primary studies assessing the diagnostic performance of PCR using DNA extracted from DBS and whole blood for detecting malarial parasites were obtained from the ISI Web of Science, Scopus, and PubMed databases. Odds ratios (ORs) and 95% confidence intervals (CIs) were plotted in forest plots using Review Manager version 5.3. Statistical analysis was performed via random-effects meta-analysis. Data heterogeneity was assessed using the I2 statistic. Of the 904 studies retrieved from the databases, seven were included in this study. The pooled meta-analysis demonstrated no significant difference in the comparative performance of PCR for detecting malaria parasites between DNA extracted from DBS and that extracted from whole blood (OR 0.85; 95% CI 0.62–1.16; I2 = 78%). However, subgroup analysis demonstrated that PCR using DNA extracted from DBS was less accurate in detecting Plasmodium vivax than that using DNA extracted from whole blood (OR = 0.85; 95% CI 0.77–0.94). In conclusion, a significant difference in detecting P. vivax was observed between PCR using DNA extracted from DBS and that using DNA extracted from whole blood. Therefore, P. vivax in endemic areas should be identified and detected with care with PCR using DNA obtained from DBS which potentially leads to a negative result. Further studies are required to investigate the performance of PCR using DBS for detecting P. vivax and other malarial parasites to provide data in research and routine surveillance of malaria, especially with renewed efforts towards the eradication of the disease.

Download Full-text

Diagnostic performance of a rapid whole blood-based RT-LAMP method for malaria diagnosis among apparently healthy blood donors and febrile neonates in Cameroon

PLoS ONE ◽

10.1371/journal.pone.0246205 ◽

2021 ◽

Vol 16 (1) ◽

pp. e0246205

Author(s):

Sylvie Georgette Zebaze Temgoua Kemleu ◽

Laure Ngando ◽

Elvige Nguekeng ◽

Balotin Fogang ◽

Marie Mafo Kapen ◽

...

Keyword(s):

Light Microscopy ◽

Whole Blood ◽

Diagnostic Performance ◽

Blood Donors ◽

Malaria Diagnosis ◽

Lamp Method ◽

Malaria Parasites ◽

Blood Samples ◽

Amplification Method ◽

Apparently Healthy

Light microscopy and rapid diagnostic tests are the two commonly used methods for malaria diagnosis that rely on the direct use of unprocessed blood samples. However, both methods do not have the level of sensitivity required for malaria diagnosis in cases of low density parasitaemia. We report here the diagnostic performance of a whole blood-based reverse transcription loop-mediated isothermal amplification method for Plasmodium falciparum malaria diagnosis in apparently healthy blood donors and febrile neonates in Cameroon. The presence of malaria parasites in whole blood samples was determined by light microscopy, antigen-based rapid diagnostic test (RDT), and by RT-LAMP using a “lyse and amplify” experimental protocol. Of the 256 blood donors tested, 36 (14.1%) were positive for malaria parasites by light microscopy, 38 (14.8%) were positive by RDT whereas 78 (30.5%) were positive by RT-LAMP. Only light microscopy and RT-LAMP detected infection among the febrile neonates (279 neonates, median age: 2 days, range: 1–9 days), with positivity rates of 8.6% and 12.2%, respectively. The overall concordance between the three methods were 75.9% for RT-LAMP and light microscopy, 75.1% for RT-LAMP and RDT, and 83.9% for light microscopy and RDT. Blood parasite densities were significantly lower in the neonates (mean: 97.6, range: 61–192 parasites/μL) compared to the blood donors (mean: 447.8, range: 63–11 000 parasites/μL). Together, the study demonstrates the usefulness of whole blood RT-LAMP for use in rapid pre-screening of blood donors and suspected neonates to avert severe consequences of P. falciparum infections.

Download Full-text

Comparison of FilmArray and Quantitative Real-Time Reverse Transcriptase PCR for Detection of Zaire Ebolavirus from Contrived and Clinical Specimens

Journal of Clinical Microbiology ◽

10.1128/jcm.01317-15 ◽

2015 ◽

Vol 53 (9) ◽

pp. 2956-2960 ◽

Cited By ~ 30

Author(s):

Timothy R. Southern ◽

Lori D. Racsa ◽

César G. Albariño ◽

Paul D. Fey ◽

Steven H. Hinrichs ◽

...

Keyword(s):

Nucleic Acids ◽

Reverse Transcriptase ◽

Real Time ◽

Whole Blood ◽

Limit Of Detection ◽

Clinical Specimens ◽

Qrt Pcr ◽

Zaire Ebolavirus ◽

Pcr Assays ◽

Blood Specimens

Rapid, reliable, and easy-to-use diagnostic assays for detection of Zaire ebolavirus (ZEBOV) are urgently needed. The goal of this study was to examine the agreement among emergency use authorization (EUA) tests for the detection of ZEBOV nucleic acids, including the BioFire FilmArray BioThreat (BT) panel, the FilmArray BT-E panel, and the NP2 and VP40 quantitative real-time reverse transcriptase (qRT) PCR assays from the Centers for Disease Control and Prevention (CDC). Specimens used in this study included whole blood spiked with inactivated ZEBOV at known titers and whole-blood, plasma, and urine clinical specimens collected from persons diagnosed with Ebola virus disease (EVD). The agreement for FilmArray and qRT-PCR results using contrived whole-blood specimens was 100% (6/6 specimens) for each ZEBOV dilution from 4 × 107to 4 × 10250% tissue culture infective dose (TCID50)/ml, as well as the no-virus negative-control sample. The limit of detection for FilmArray and qRT-PCR assays with inactivated ZEBOV, based on duplicate positive results, was determined to be 4 × 102TCID50/ml. Rates of agreement between FilmArray and qRT-PCR results for clinical specimens from patients with EVD were 85% (23/27 specimens) for whole-blood specimens, 90% (18/20 specimens) for whole-blood specimens tested by FilmArray testing and matched plasma specimens tested by qRT-PCR testing, and 85% (11/13 specimens) for urine specimens. Among 60 specimens, eight discordant results were noted, with ZEBOV nucleic acids being detected only by FilmArray testing in four specimens and only by qRT-PCR testing in the remaining four specimens. These findings demonstrate that the rapid and easy-to-use FilmArray panels are effective tests for evaluating patients with EVD.

Download Full-text

Quantitative Detection of Plasmodium falciparum Using, LUNA-FL, A Fluorescent Cell Counter

Microorganisms ◽

10.3390/microorganisms8091356 ◽

2020 ◽

Vol 8 (9) ◽

pp. 1356 ◽

Cited By ~ 1

Author(s):

Muneaki Hashimoto ◽

Kazumichi Yokota ◽

Kazuaki Kajimoto ◽

Musashi Matsumoto ◽

Atsuro Tatsumi ◽

...

Keyword(s):

Whole Blood ◽

Point Of Care ◽

Regression Line ◽

Malaria Diagnosis ◽

Quantitative Detection ◽

Fluorescent Cell ◽

Blood Samples ◽

Parasite Detection ◽

Whole Blood Samples ◽

Cell Counter

The microscopic examination of Giemsa-stained thin and/or thick blood films (Giemsa microscopy) is the standard method of malaria diagnosis. However, the results of the diagnosis significantly depend on the skills of clinical technicians. Furthermore, sample preparation and analysis are laborious and time-consuming. Therefore, in this study, we investigated if a commercially available fluorescent cell counter, LUNA-FL, was useful for the detection of Plasmodium parasite and the estimation of parasitemia. Whole blood samples from uninfected persons, spiked with P. falciparum-infected erythrocytes, were analysed. Most of the leucocytes and platelets were removed from whole blood samples with SiO2-nanofiber filters set on spin columns. The filtered samples were stained with acridine orange, and automatic detection, as well as counting of erythrocytes and parasites, were performed using LUNA-FL. Whole blood, with various levels of parasites, was analysed by Giemsa microscopy or with LUNA-FL to estimate parasitemia, and a comparative analysis was performed. The coefficient determination value of the regression line was high (R2 = 0.98), indicating that accurate quantitative parasite detection could be performed using LUNA-FL. LUNA-FL has a low running cost; it is compact, fast, and easy to operate, and may therefore be useful for point-of-care testing in the endemic areas.

Download Full-text

Portable optofluidic absorption flow analyzer for quantitative malaria diagnosis from whole blood

Applied Optics ◽

10.1364/ao.55.008637 ◽

2016 ◽

Vol 55 (30) ◽

pp. 8637 ◽

Cited By ~ 4

Author(s):

Earu Banoth ◽

Vamshi Krishna Kasula ◽

Sai Siva Gorthi

Keyword(s):

Whole Blood ◽

Malaria Diagnosis

Download Full-text

Fine Structure of Platelet Interaction with a New Microcrystalline Collagen Preparation

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100050305 ◽

1974 ◽

Vol 32 ◽

pp. 58-59

Author(s):

W. H. Zucker ◽

R. G. Mason

Keyword(s):

Fine Structure ◽

Platelet Aggregation ◽

Hydrochloric Acid ◽

Whole Blood ◽

Platelet Adhesion ◽

Hemostatic Agent ◽

Native Collagen ◽

Fibrillar Morphology ◽

Clinical Interest ◽

New Form

Platelet adhesion initiates platelet aggregation and is an important component of the hemostatic process. Since the development of a new form of collagen as a topical hemostatic agent is of both basic and clinical interest, an ultrastructural and hematologic study of the interaction of platelets with the microcrystalline collagen preparation was undertaken.In this study, whole blood anticoagulated with EDTA was used in order to inhibit aggregation and permit study of platelet adhesion to collagen as an isolated event. The microcrystalline collagen was prepared from bovine dermal corium; milling was with sharp blades. The preparation consists of partial hydrochloric acid amine collagen salts and retains much of the fibrillar morphology of native collagen.

Download Full-text

3-D organization of fibrinogen receptors using computer reconstruction and whole-mount microscopy

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100102237 ◽

1988 ◽

Vol 46 ◽

pp. 30-31

Author(s):

E. T. O'Toole ◽

R. R. Hantgan ◽

J. C. Lewis

Keyword(s):

Whole Blood ◽

Colloidal Gold ◽

Blood Platelets ◽

Cell Contact ◽

Computer Assisted ◽

Adherent Cells ◽

Differential Centrifugation ◽

Computer Reconstruction ◽

Sds Page ◽

Whole Mount

Thrombocytes (TC), the avian equivalent of blood platelets, support hemostasis by aggregating at sites of injury. Studies in our lab suggested that fibrinogen (fib) is a requisite cofactor for TC aggregation but operates by an undefined mechanism. To study the interaction of fib with TC and to identify fib receptors on cells, fib was purified from pigeon plasma, conjugated to colloidal gold and used both to facilitate aggregation and as a receptor probe. Described is the application of computer assisted reconstruction and stereo whole mount microscopy to visualize the 3-D organization of fib receptors at sites of cell contact in TC aggregates and on adherent cells.Pigeon TC were obtained from citrated whole blood by differential centrifugation, washed with Ca++ free Hank's balanced salts containing 0.3% EDTA (pH 6.5) and resuspended in Ca++ free Hank's. Pigeon fib was isolated by precipitation with PEG-1000 and the purity assessed by SDS-PAGE. Fib was conjugated to 25nm colloidal gold by vortexing and the conjugates used as the ligand to identify fib receptors.

Download Full-text