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Query: UMLS:C0024530 (malaria)
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This study evaluated a newly developed rapid malaria diagnostic test, OptiMAL Assay, to detect "Plasmodium falciparum malaria" and "non Plasmodium falciparum malaria" in blood samples from 139 individuals with a presumptive clinical diagnosis of imported malaria in Italy. OptiMAL Assay utilizes a dipstick coated with monoclonal antibodies against the intracellular metabolic enzyme, plasmodium Lactate Dehydrogenase (pLDH) present in and released from parasite-infected erythrocytes. Blood samples from 56 cases out of 139 were found "Plasmodium falciparum malaria" positive by microscopy; with these samples OptiMAL Assay and the ParaSight-F test, which is a kit detecting the P. falciparum histidin-rich protein 2 (HRP-2), showed an overall sensitivity of 83% and 94%, respectively, in comparison with microscopy. Parasitemia levels tested in the 56 P. falciparum positive blood samples by microscopy ranged from <0.004% to 20%. A correlation between sensitivity and parasitemia was evident and OptiMAL Assay and ParaSight-F test were more sensitive (96-100%; 100%) with samples with 0.1%-20% levels of parasitemia, while proved less sensitive (0-44%; 50-88%) with <0.004-0.01% levels of parasitemia.
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PMID:Evaluation of OptiMAL Assay test to detect imported malaria in Italy. 1106 27

Malaria is one of the most important parasitic diseases especially in tropical areas. Over 300 million people are affected and the condition causes 1-3 million deaths each year. It is transmitted by the bite of infected Anopheles mosquitoes. Although Korea was declared to be free of Malaria by the WHO in 1979, malaria re-emergence has been apparent since 1993 amongst soldiers located near the De-Militarized Zone (DMZ) in the northern part of the country. Conventional microscopic examination of thin and thick blood films demonstrates the presence of the parasite and thus this method has been used to confirm the diagnosis of malaria, but it is a labor-intensive procedure and relies upon subjective interpretation. To overcome these limitations, fast and reliable methods for malaria detection have been recently introduced. In this study, we compared three kinds of antibody detection kits and one biochemical test kit that determines the presence of Plasmodium lactate dehydrogenase (pLDH) with conventional peripheral blood smears. The antibody detection methods examined were, two rapid test pack format methods and a single microplate format enzyme-linked immunosorbent assay (ELISA) kit, as manufactured by Korean companies. The sensitivities of the three commercial antibody detection kits in the early stage of malaria were 70.8%, 77.4%, and 63.6%, their corresponding specificities 90.5%, 91.8%, and 80.9%, and their accuracies 87.6%, 87.0%, and 76.7%. The sensitivity and specificity of the pLDH assay were 100% apiece and the results were in 100% concordance with the microscopy of thick blood films. Thus, the pLDH assay may be used as an alternative for conventional microscopic blood film examination, especially in emergency situations when prompt treatment is necessary.
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PMID:Evaluation of diagnostic methods of re-emerging malaria in Korean patients. 1129 5

Recombinant glutathione S-transferase (agGST1-6) from the malaria vector mosquito Anopheles gambiae Giles (Diptera: Culicidae) was expressed in Escherichia coli using a pET3a vector system. The expressed enzyme was biochemically active with reduced glutathione (GSH) and 1-chloro-2,4-dinitrobenzene (CDNB). Activity of agGST1-6 with GSH and CDNB was inhibited to different degrees by both alpha-cyano and non-alpha-cyano pyrethroid insecticides. This inhibition was used to develop an assay for quantification of pyrethroids. Standard curves of insecticide concentration against percentage of enzyme inhibition or volume of iodine solution were established by spectrophotometry and iodine volumetric titration, respectively, for permethrin and deltamethrin. These assays allowed estimation of pyrethroid concentrations both spectrophotometrically and visually. For the residue assay of each insecticide, a cut-off point of 50% of the initial pyrethroid impregnation concentration was used, which should differentiate between biologically active and inactive treated bednets. The cross-reactivity of the primary permethrin photodegradants (3-phenoxyalcohol and 3-phenoxybenzoic acid) with the recombinant agGST1-6 was assayed in the same system. No agGST1-6 inhibition by the insecticide metabolites was observed, suggesting that the system is unaffected by primary permethrin metabolites and will accurately measure insecticide parent compound concentrations. The estimated pyrethroid insecticide concentrations, given spectrophotometrically and by iodine titration assay, were comparable to those obtained by direct HPLC quantification of residues extracted from bednets. Hence, it should be relatively easy to adapt this method to produce a test kit for residue quantification in the field.
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PMID:Quantification of pyrethroid insecticides from treated bednets using a mosquito recombinant glutathione S-transferase. 1129 2

Lack of experience on the part of involved laboratory personnel frequently complicates swift diagnosis of imported falciparum malaria in non-endemic areas. Diagnostic tools based on the dipstick principle for the detection of plasmodial histidine-rich protein 2 have been marketed for several years and have been extensively evaluated. Recently, a test kit capable of detecting antigen of Plasmodium falciparum and P. vivax has been introduced. In order to evaluate this newly available tool, specimens from 664 patients were screened during the course of a prospective multicentre study within the European Network on Imported Infectious Disease Surveillance (TropNetEurop). Among the screened specimens, samples from 82 patients (12.3%) were positive for falciparum malaria using expert microscopy. A further 17 samples were positive for vivax malaria. The evaluated test kit performed with a sensitivity of 87.8% and a specificity of 99% for detection of falciparum malaria. Respective values for vivax malaria were 76.5% and 100%. Dipstick tests have the potential of improving the speed and accuracy of the diagnosis of falciparum malaria, especially if non-specialized laboratories are involved. However, decreased values of sensitivity and specificity, in comparison with expert microscopy, still impose a clear limit on the usefulness of the currently available kits.
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PMID:Evaluation of a dipstick test for the rapid diagnosis of imported malaria among patients presenting within the network TropNetEurop. 1172 42

Logistic, economic and technical factors limit rapid access to microscopic confirmation of symptomatic diagnosis of malaria in many rural areas in endemic countries such as Myanmar. A study was conducted to evaluate a rapid on-site immunochromatographic test (ICT Malaria Pf/Pv) for detection of Plasmodium falciparum and P. vivax in two villages in the Taikkyi region of Myanmar. The ICT Malaria tests were performed by a volunteer health worker (VHW) in Yae-Aye-San village and by a professionally trained midwife (MW) in Kankone village. A total of 1000 symptomatic patients participated in the study by providing blood samples for an ICT test and for microscopy. The ICT performance indices, relative to microscopy, were better for the trained MW compared with the less experienced VHW. For P. falciparum and/or P. vivax infections, the sensitivities were 82.7% for the VHW compared with 93.7% for the MW. For P. falciparum infections, the sensitivities were 82.2% for the VHW and 91.3% for the MW, while the corresponding values for P. vivax infections were 66.7 and 79%, respectively. Although the test kit appeared to perform better in more experienced hands, this study questions whether this difference is related to the use of the ICT Malaria Pf/Pv test kit, or related to other factors such as differences in the quality of blood slides prepared by the VHW and MW for microscopic examination. Overall, the results suggest that a rapid diagnostic assay such as the ICT Malaria Pf/Pv test kit can be used in rural settings by relatively inexperienced persons, such as VHWs, with a reasonable degree of sensitivity, thus providing on-site confirmation of symptomatic diagnosis of malaria.
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PMID:Performance appraisal of rapid on-site malaria diagnosis (ICT malaria Pf/Pv test) in relation to human resources at village level in Myanmar. 1175 28

Blood samples collected from 34 patients with severe malaria who were involved in antimalarial treatment studies were tested with rapid dipstick assay (Rapid Test Malaria, RTM from Quorum Diagnostics Inc., Vancouver, BC, Canada), based on the detection of Histidine Rich Protein (HRP-2) of Plasmodium falciparum. This was compared with the conventional Giemsa stained thin and thick blood smears. The study was done from March 1998 to May 1998, at the Basic Research Laboratory of the Faculty of Medicine, Addis Ababa University. Comparable number of patients (n = 32) with various diagnosis other than falciparum malaria were used as controls. The rapid dip-stick assay was positive in 31 among 34 of the severe malaria cases with sensitivity of 91.2%, specificity of 93.7%, positive predictive value (PPV) of 93.9% and negative predictive value (NPV) of 90.9%. The three cases missed by the RTM, had parasitemia of 66,000, 44,000, and 40,000/microL of blood which might be due to genetic heterogeneity of the HPR-2 expression. Among the controls, there were 2 false positive cases which may be as a result of persistent HPR-2 antigen after the clearance of peripheral parasitemia. The dip-stick method is a very quick, sensitive and specific diagnostic tool with limits of detection comparable or better than those provided by the light microscopy. The simplicity of the technique makes this method more applicable in the resource deprived laboratories of developing countries provided the kit is affordable for large scale use.
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PMID:Rapid diagnosis of severe malaria based on the detection of Pf-Hrp-2 antigen. 1195 14

We investigated the prevalence of infection with hepatitis B virus among adult Vietnamese patients hospitalized for severe Plasmodiumfalciparum malaria. Sera from patients admitted with severe malaria in Ho Chi Minh City, Vietnam, between May 1991 and January 1996 were assayed for hepatitis B surface antigen (HB(s)Ag) by a commercial enzyme-linked immunosorbent assay kit. The overall prevalence of HB(s)Ag was 23.77% (77 of 324). This was higher than reported estimates of prevalence in the general catchment population for the study hospital (mean, 9.8%; range, 9-16%). No association was found between risk of death caused by severe malaria and HB(s)Ag. Patients admitted with cerebral malaria had a slightly greater risk of registering positive for HB(s)Ag (relative risk, 1.28; 95% confidence interval, 1.04-1.58) relative to other manifestations of severe malaria. Chronic infection with hepatitis B virus may be a risk factor for severe malaria.
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PMID:Short report: hepatitis b infection and severe Plasmodium falciparum malaria in Vietnamese adults. 1213 83

Circumsporozoite (CS) antibodies are a reliable serological marker for the infection of Plasmodium falciparum. The purpose of this investigation was to construct and evaluate an enzyme-linked immunosorbent assay test for the detection of CS antibodies. While the sensitivity of the newly developed test reached 78%, the specificity was 99%. In addition, the optimized kit was used to test for infection with P. falciparum in 1903 travellers that were recruited from a prospective study for malaria chemoprophylaxis. Sixty-six of the 1903 patients (3.5%) showed elevated CS antigen antibody titres. However, seroconversion could only be demonstrated in 18 (0.95%) patients. Among those seroconverting, there was a significantly higher percentage of male travellers (1.28%) than female travellers (0.56%). Positive reactions were more frequent among returnees from West and East Africa (1.49 and 1.14%, respectively) than among those from other endemic areas, e.g. South America (n=0). Despite its limited sensitivity, this newly developed kit for CS antibody testing may be a valuable tool for the estimation of the risk for travellers in malarious regions to acquire an infection with P. falciparum. It may also be useful for the determination of the efficacy of malaria chemoprophylaxis for inhibiting outbreak of disease.
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PMID:Plasmodium falciparum: use of a NANP19 antibody-test for the detection of infection in non-immune travellers. 1216 92

Many types of ELISA-based immunodiagnostic test kits are commercially available in the market for specific indications. These kits provide necessary assay components, reagents, and guidelines to perform the assay under designated optimal conditions. By using these kits, any unknown or test sample can be assessed as negative or positive based on the results of referral calibrator (Ref+ve and Ref-ve) samples. It is essential to provide reliable test kits to end-users with adequate quality control analysis. Therefore, it is necessary to check the kit for any variations in its performance. While developing a malaria antibody ELISA test-kit, we optimized assay conditions with chequer-board analyses and developed an assay protocol. We have taken out kits randomly from the assembly line and had them evaluated by operators who are new to the test-kits. Assays are performed as per the test guidelines provided. Sera, diluted serially, have shown a clear discriminatory signal between a negative vs. positive sample. A COV is determined by evaluating the Ref-ve calibrator in replicate antigen-coated wells from 6 different plates. This COV is used as a tool to determine S/N ratio of test samples. Besides Ref-ve and Ref+ve calibrators, additional field serum samples are tested with the test kit. Several performance indices, such as mean, standard deviation, %CV are calculated, and the inter- and intra-assay variations determined. The assay precision is determined with large and small replicate samples. In addition, assays are performed concurrently in triplicate-, duplicate-, and single-wells, and the results are analyzed for any assay variations. Different plate areas are identified in antigen-coated 96-well plates and tested blind to detect any variations. The S/N ratio is found to be a very effective tool in determining the assay sensitivity. The %CV was within 10-15%. Variations seen in the assays are found to be due to operator errors and not due to kit reagents. These observations, although, are based upon one type; however, it may as well apply to other line of kits. This is obviously valuable to the end-users of ELISA kits. The operator related error has to be ascertained before lodging any complaint on the kit performance. Based on this data, the test kit has shown acceptable sensitivity and precision and offers compliance on the way the test kits is manufactured. With this, it is concluded that the test kits are suitable for detecting malaria antibody in clinical sample analysis.
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PMID:Assessment of assay sensitivity and precision in a malaria antibody ELISA. 1268 Jun 9

Malaria is a major parasitic disease in tropical areas. Three to five hundred million people suffer from the disease and it kill a million people per year. Blood smear observation was developed for the diagnosis of malaria, but the examination needs skilled experts and exact diagnosis is time consuming. A kit based on immunochromatography can be a reliable and rapid method for clinical diagnosis, even in the hands of inexperienced personnel. However, all such currently developed kits can only diagnose P. falciparum malaria. In our previous report, the C-terminal region of P. vivax merozoite surface protein 1 (PvcMSP) was cloned and expressed in E. coli. In the present study, we developed an immunochromatographic kit using this PvcMSP for the diagnosis of specific antibody to P. vivax malaria in serum samples. The kit was used to examine sera from vivax malaria patients and non-malaria- infected person and the test showed 100% sensitivity (78/78) and 98.3% specificity (58/59). This result demonstrated that the immunochromatographic kit for P. vivax antibody detection is applicable for the rapid and precise diagnosis of P. vivax malaria.
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PMID:Development and evaluation of an immunochromatographic kit for the detection of antibody to Plasmodium vivax infection in South Korea. 1295 Jan 38


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