Detection of Plasmodium vivax and Plasmodium falciparum DNA in human saliva and urine: Loop-mediated isothermal amplification for malaria diagnosis

This study investigated loop-mediated isothermal amplification (LAMP) detection of Plasmodium falciparum and Plasmodium vivax in urine and saliva of malaria patients. From May to November 2011, 108 febrile patients referred to health centers in Sistan and Baluchestan Province of south-eastern Iran participated in the study. Saliva, urine, and blood samples were analyzed with nested PCR and LAMP targeting the species-specific nucleotide sequence of small subunit ribosomal RNA gene (18S rRNA) of P. falciparum and P. vivax and evaluated for diagnostic accuracy by comparison to blood nested PCR assay. When nested PCR of blood is used as standard, microscopy and nested PCR of saliva and urine samples showed sensitivity of 97.2%, 89.4% and 71% and specificity of 100%, 97.3% and 100%, respectively. LAMP sensitivity of blood, saliva, and urine was 95.8%, 47% and 29%, respectively, whereas LAMP specificity of these samples was 100%. Microscopy and nested PCR of saliva and LAMP of blood were comparable to nested PCR of blood (к = 0.95, 0.83, and 0.94, respectively), but agreement for nested PCR of urine was moderate (к = 0.64) and poor to fair for saliva LAMP and urine LAMP (к = 0.38 and 0.23, respectively). LAMP assay showed low sensitivity for detection of Plasmodium DNA in human saliva and urine compared to results with blood and to nested PCR of blood, saliva, and urine. However, considering the advantages of LAMP technology and of saliva and urine sampling, further research into the method is worthwhile. LAMP protocol and precise preparation protocols need to be defined and optimized for template DNA of saliva and urine.     

Changing trends in prevalence of different Plasmodium species with dominance of Plasmodium falciparum malaria infection in Aligarh (India)

ObjectiveTo determine the prevalence of malaria in Aligarh and analyze species dominance in different years over a decade.MethodsDiagnosis of malaria was done using microscopy as gold standard, rapid antigen detection assays and quantitative buffy coat (QBC) assays. Giemsa stained blood smear examination was done, thick and thin films were examined for presence of different Plasmodium spp. Rapid antigen detection assays employing detection of HRP-2 and parasite lactate dehydrogenase antigen (pLDH) by immunochromatography was done in patients whose blood smear found to be negative by conventional Giemsa slide examination. QBC was done in cases where there is strong clinical suspicion of malaria with blood smear negative, in patients with chronic malaria, splenomegaly, or in those patients who had inadequate treatment and for post-treatment follow up.ResultsPlasmodium vivax and Plasmodium falciparum were only species detected in our hospital. Overall prevalence of malaria in Aligarh was found to be 8.8%. The maximum prevalence of 20.1% was observed in year 2008 and lowest 2.3% in 2002.ConclusionsHigh prevalence of malaria is observed in this part of country with dominance of both species particularly Plasmodium falciparum should be monitored and factors accounting for occurrence should be studied to employ effective control measures.     

Accuracy of Two Malaria Rapid Diagnostic Tests (RDTS) for Initial Diagnosis and Treatment Monitoring in a High Transmission Setting in Uganda

Malaria rapid diagnostic tests (RDTs) may improve fever management in areas without microscopy. We compared the accuracy of histidine-rich protein 2 (HRP2) and Plasmodium lactate dehydrogenase (pLDH)-based RDTs, using expert microscopy as a gold standard, for initial diagnosis, treatment monitoring, and diagnosis of recurrent malaria in a cohort of children followed longitudinally in a high-transmission area in Uganda. For 305 initial fever episodes, sensitivity was 98% for HRP2 and 87% for pLDH, whereas specificity was 55% and 96%, respectively. The HRP2 gave 51% false-positive results on Day 28, whereas pLDH gave no false positives after Day 7. For 59 recurrent fever episodes during follow-up, sensitivity was 100% for HRP2 and 91% for pLDH, whereas specificity was 33% and 100%, respectively. The HRP2-based RDTs are useful for initial diagnosis of malaria caused by superior sensitivity; however, as a result of superior specificity, pLDH-based RDTs are more appropriate to monitor treatment and diagnose recurrent malaria.     

Parasite-specific lactate dehydrogenase for the diagnosis of Plasmodium falciparum infection in an endemic area in West Uganda

The measurement of parasite lactate dehydrogenase (pLDH) has been presented as an easy and rapid method for the diagnosis of malaria in humans. In order to evaluate the sensitivity and specificity of such a test we examined blood samples from 429 Ugandan patients. While pLDH activity was significantly linked to parasitaemia, sensitivity and specificity were found to be rather low at 58.8 and 62.2% respectively. The positive and negative predictive values failed to meet necessary standards. We conclude that the methods of measurement of pLDH activity in malaria infection, although potentially useful for the fast diagnosis of malaria, need to be improved to be of true value in endemic areas.     

Field evaluation of rapid diagnostic tests for malaria in Yaounde,Cameroon

Rapid diagnostic tests (RDTs) are affordable, alternative diagnostic tools. The present study aimed to evaluate RDTs available in Cameroon and compare their characteristics to follow the parasitological response of patients for 28 days. Malaria diagnosis was assessed in 179 febrile patients using conventional microscopy as the reference method. Parascreen detects both Plasmodium falciparum-specific histidine-rich protein 2 (Pf HRP-2) and Pan-specific plasmodial lactate dehydrogenase (pLDH) in all four human Plasmodium spp. Diaspot is based on the detection of Pf HRP-2. OptiMAL-IT (pLDH specific for P. falciparum and pLDH for all four human Plasmodium spp.) was assessed for comparison. The reliability of RDTs was evaluated by calculating the sensitivity, specificity, positive predictive value, negative predictive value, false-positive rate, false-negative rate, and likelihood ratio. The clinical outcome of 18 children treated with atovaquone–proguanil and followed for 28 days was evaluated using microscopy and RDTs. Of 179 samples, 133 (74.3%) were pure P. falciparum-positive smears, 4 (2.2%) pure P. malariae-positive smears, and 42 (23.5%) negative smears. Parascreen and Diaspot had high sensitivity (>92%) and positive predictive values (>94%). The specificities for Parascreen and Diaspot were 81.0% and 90.5%, respectively. The false-positive rates and the false-negative rates were 19.0% and 4.5% for Parascreen and 9.5% and 8.3% for Diaspot, respectively. Most false-negatives occurred in samples with low parasitaemia (<500 asexual parasites/μL). The performance of RDTs was better at higher parasitaemia (>500 asexual parasites/μL). Four pure P. malariae were only detected by the pan-Plasmodium bands of Parascreen and OptiMAL-IT. In blood samples from patients treated and followed-up for 28 days, HRP2-based RDTs remained positive in most samples until Day 28. Despite negative smears, OptiMAL-IT remained positive in several patients until Day 7 but was negative in all patients from Day 14 onwards. RDTs can improve the management of febrile patients. The validity, ease of use, and cost of HRP2-based tests were comparable. However, one of the current weaknesses of the RDT-based strategy using the tests available in Cameroon is inadequate sensitivity for low parasitaemia. In some cases, RDT results may require correct interpretation based on clinical history, clinical examination, and microscopic diagnosis.     

Automated detection of malaria‐associated intraleucocytic haemozoin by Cell‐Dyn CD4000 depolarization analysis

Summary Laboratory tests for malaria are only performed if there is clinical suspicion of the disease, and a missed diagnosis contributes substantially to morbidity and mortality. Malaria parasites produce haemozoin, which is able to depolarize light and this allows the automated detection of malaria during routine complete blood count analysis (CBC) with some Abbott Cell‐Dyn instruments. In this study, we evaluated the Cell‐Dyn CD4000 with 831 blood samples submitted for malaria investigations. Samples were categorized as malaria negative (n = 417), convalescent malaria (n = 64) or malaria positive (n = 350) by reference to thin/thick film microscopy, ‘rapid test’ procedures, polymerase chain reaction analysis and clinical history. With regard to CD4000 depolarization analysis, a malaria positive CD4000 pattern was ascribed to samples that showed one or more abnormal depolarizing purple events, which corresponded to monocytes containing ingested malaria pigment (haemozoin). Positive CD4000 patterns were observed in 11 of 417, 50 of 64 and 281 of 350 of malaria negative, convalescent malaria and malaria positive samples respectively. The specificity and positive predictive values for malaria (active and convalescent) were very high (97.4 and 96.8%, respectively), while sensitivity and negative predictive values were 80.0 and 83.0% respectively. Depolarization analysis was particularly effective for Plasmodium falciparum malaria but there was lower detection sensitivity for White compared with Black African patients. CD4000 90° depolarization vs 0° analysis revealed a proportion of samples with small nonleucocyte‐associated depolarizing particles. Appearance of such events in the form of a discrete cluster was associated with P. vivax rather than P. falciparum infection.     

Comparative Diagnosis of Malaria Infections by Microscopy,Nested PCR,and LAMP in Northern Thailand

Three methods, microscopy, nested polymerase chain reaction (nPCR), and loop-mediated isothermal amplification (LAMP) have been applied for malaria diagnosis in 105 human blood samples collected in Northern Thailand. Only Plasmodium falciparum and Plasmodium vivax infections were detected. A total number of 57 positives (54%) could be detected for P. falciparum and 25 (24%) for P. vivax when all samples that were positive in any of the three methods are counted together. The nPCR was used as a reference standard for comparison with the other methods, microscopy and LAMP. The sensitivity of LAMP for P. falciparum was 100%. All nPCR-negative samples for P. falciparum were also negative by both microscopy and LAMP (specificity, 100%). For diagnosis of P. vivax, microscopy detected 15 of 23 nPCR-positive samples (sensitivity, 65%). LAMP detected 22 of 23 nPCR-positives (sensitivity, 96%). Among the 82 nPCR-negative samples microscopy detected two samples (specificity, 98%). All 82 nPCR-negative were also negative by the LAMP method (specificity, 100%). Both Plasmodium genus- and species-specific LAMP primer sets yielded the same results in all samples. There were no significant differences in the prevalence detected by each method. We assume that LAMP was as reliable as nPCR and more reliable than microscopy in the detection of Plasmodium DNA tested in the examined Thai field blood samples. This study further validates LAMP as an alternative molecular diagnostic tool, which can be used in the diagnosis of early infections of malaria cases and together with nPCR can also be used as supplementary methods for clinical and epidemiological use.     

Comparative evaluation of microscopy,OptiMAL® and 18S rRNA gene based multiplex PCR for detection of Plasmodium falciparum & Plasmodium vivax from field isolates of Bikaner,India

ObjectiveTo evaluate microscopy, OptiMAL® and multiplex PCR for the identification of Plasmodium falciparumm (P. falciparum) and Plasmodium vivax (P. vivax) from the field isolates of Bikaner, Rajasthan (Northwest India).MethodsIn this study, a multiplex PCR (P. falciparum and P. vivax) was further developed with the incorporation of Plasmodium malariae (P. malariae) specific primer and also a positive control. The performance of microscopy, plasmodium lactate dehydrogenase (pLDH) based malaria rapid diagnostic test OptiMAL® and 18S rRNA gene based multiplex PCR for the diagnosis of P. falciparum and P. vivax was compared.ResultsThe three species multiplex PCR (P. falciparum, P. vivax and P. malariae) with an inbuilt positive control was developed and evaluated. In comparison with multiplex PCR, which showed the sensitivity and specificity of 99.36% (95% CI, 98.11%–100.00%) and 100.00% (95% CI, 100.00%–100.00%), the sensitivity and specificity of microscopy was 90.44% (95% CI, 88.84%–95.04%) and 99.22% (95% CI, 97.71%–100.00%), and OptiMAL® was 93.58% (95% CI, 89.75%–97.42%) and 97.69% (95% CI, 95.10%–100.00%). The efficiencies were 99.65%, 95.10% and 95.45% for multiplex PCR, microscopy and OptiMAL®, respectively.ConclusionsOur results raise concerns over the overall sensitivities of microscopy and OptiMAL®, when compared to the multiplex PCR and thus stress the need for new molecular interventions in the accurate detection of the malarial parasites. This further highlights the fact that further developments are needed to improve the performance of rapid diagnostic tests at field level.     

Comparison of Paracheck Pf® test with conventional light microscopy for the diagnosis of malaria in Ethiopia

ObjectiveTo assess the accuracy of Paracheck Pf® in reference to the conventional light microscopy.MethodsA total of 400 patients visiting Awash, Methara and Ziway malaria centers were simultaneously screened with both light microscopy and Paracheck Pf® for the presence of Plasmodium falciparum (P. falciparum) malaria.ResultsOf the 190 samples that were negative by light microscope, the Paracheck Pf® showed 11 false positive and 179 true negative results, and from a total of 210 samples positive by light microscope, Paracheck Pf® accurately diagnosed 200 true malaria cases. Taking the light microscopy as a standard test for malaria, the sensitivity, specificity, positive predictive value and negative predictive value of Paracheck Pf® is 95.2% [cofidence interval (CI)=92.4–97.1], 94.2% (CI=91.1–96.3), 94.8% (CI=92.0–96.7) and 94.7% (CI=91.6–96.8), respectively.ConclusionsParacheck Pf® showed good sensitivity and specificity for the diagnosis of P. falciparum malaria, and fulfill the world health organization (WHO) recommendation that requires the sensitivity of rapid diagnostic tests (RDTs) to be greater than 95%. Therefore, Paracheck Pf® can be used as an alternative to the Giemsa stain light microscopy in resource poor set ups.     

试条法快速诊断恶性疟的研究

目的 :探讨试条法检测恶性疟患者血液中富组氨酸蛋白 快速诊断恶性疟在海南疟区现场的诊断价值。方法 :用试条法检测 66例发热病人血样 ,将结果同血涂片镜检法进行比较。结果 :试条法检出恶性疟 (包括混合感染 )的敏感度和特异度分别为 85.7%和95.6% ,两种检测法的符合率为 92 .4 % ;用试条法检测 2 0例间日疟患者血样无交叉反应。结论 :试条法诊断恶性疟快速方便 ,能诊断早期恶性疟患者。     

Development of a Polymerase Chain Reaction (PCR) method based on amplification of mitochondrial DNA to detect Plasmodium falciparum and Plasmodium vivax

In this study we standardized a new technical approach in which the target mitochondrial DNA sequence (mtDNA) is amplified using a simple but sensitive PCR method as a tool to detect Plasmodium falciparum and Plasmodium vivax. Specific primers were designed to hybridize with cytochrome c oxidase genes of P. falciparum (cox III) and P. vivax (cox I). Amplification products were obtained for all positive samples, presenting homology only for species-specific mtDNA. Sensitivity and specificity were 100%. The applicability of the method was tested in a cross-sectional study, in which 88 blood samples from individuals naturally exposed to malaria in the Brazilian Amazon region were analyzed. Based on the results, the sensitivity and specificity were 100% and 88.3%, respectively. This simple and sensitive PCR method can be useful in specific situations and in different settings of malaria management, in endemic as well as non-endemic areas (travelers), and in clinical or epidemiological studies, with applications in malaria control programs.     

PCR扩增环子孢子蛋白基因3’ 端片段用于检测恶性疟原虫的初步研究

目的 :建立 PCR检测恶性疟原虫的新方法。方法 :作者采用自行设计并合成的一对恶性疟原虫特异引物 ,经 PCR扩增环子孢子蛋白 ( CSP)基因 3 端保守区序列 2 4 5bp片段 ,观察了它的特异性、敏感性和稳定性。结果 :1从 4株培养的恶性疟原虫和 2例恶性疟患者血样中均扩增出约 2 4 5bp的 DNA目的片段 ,用业已鉴定的 CSP基因序列作模板再行扩增证实其为恶性疟原虫CSP基因片段 ;2对间日疟原虫、利什曼原虫、弓形虫和健康人血样进行 PCR反应 ,均未见扩增条带 ;3本检测系统可检出恶性疟原虫感染血样中 0 .18个原虫所含的 DNA模板 ;4采用不同方法制备模板及不同反应方式均能获得满意结果 ;结论 :PCR扩增恶性疟原虫 CSP基因 3端片段用于恶性疟原虫检测具有灵敏、高度特异且稳定性好等优点。     

Performance of the OptiMAL assay for detection and identification of malaria infections in asymptomatic residents of Irian Jaya, Indonesia

The OptiMAL assay, a new immunochromatographic "dipstick" test for malaria based on detection of Plasmodium lactate dehydrogenase (pLDH), is purported to detect infections of approximately 200 parasites/microL of blood and to differentiate between Plasmodium falciparum and non-P. falciparum. We evaluated OptiMAL performance by comparing the test strip interpretations of two independent readers with consensus results obtained independently by expert malaria microscopists. Unbiased measures of sensitivity were derived by applying the OptiMAL test for detection and differentiation of light, asymptomatic infections by P. falciparum and Plasmodium vivax. OptiMAL readings were separated in time to determine whether the reaction signal was stable. Microscopy identified infections in 225 of 505 individuals screened; those with P. falciparum (n = 170) averaged 354 asexual forms/microL and P. vivax/Plasmodium malariae (n = 112) averaged 216 asexual forms/microL of blood. Concordance between OptiMAL and microscopy was 81% and 78% by the two independent readings. The assay's sensitivity for detection of any malaria species was 60.4% and 70.2% respectively and specificity was 97% and 89%. Most cases identified by microscopy as P. falciparum were graded as negative or non-falciparum by both OptiMAL readers. OptiMAL false negatives as well as misidentifications were related to low parasitemias (< 500/microL). The OptiMAL assay demonstrated 88-92% sensitivity for detecting infections of 500-1,000 parasites/microL, a range covering the mean parasitemia of primary symptomatic P. falciparum infections in malaria-na?ve Indonesian transmigrants. This device was markedly less sensitive than expert microscopy for discriminating between malaria species and is presently unsuited for use as an epidemiological screening tool. The OptiMAL assay is not approved for diagnostic use but is commercially available for research purposes only.     

A comparison of two rapid field immunochromatographic tests to expert microscopy in the diagnosis of malaria

In Myanmar, we tested two rapid malaria immunochromatographic kits: the OptiMAL assay for the detection of parasite lactate dehydrogenase (pLDH), and the ICT Malaria P.f./P.v. test for histidine-rich protein 2 (PfHRP2) and panmalarial antigens. A total of 229 patients were examined, of whom 133 were found to be malaria positive by Giemsa microscopy. Both OptiMAL and ICT gave lower sensitivities than previously reported. ICT sensitivity for Plasmodium falciparum and non-falciparum parasites were 86.2 and 2.9%, respectively; specificity was 76.9 and 100%, respectively. OptiMAL sensitivity for P. falciparum and non-falciparum parasites were 42.6 and 47.1%, respectively; specificity was 97.0 and 96.9%, respectively. The sensitivity of both tests for the detection of both P. falciparum and non-falciparum parasites increased with parasite density. Several explanations for these results are explored. Our results raise particular concern over batch quality variations of malaria rapid diagnostic devices (MRDDs).     

Malaria PCR Detection in Cambodian Low-Transmission Settings: Dried Blood Spots versus Venous Blood Samples

In the context of malaria elimination, novel strategies for detecting very low malaria parasite densities in asymptomatic individuals are needed. One of the major limitations of the malaria parasite detection methods is the volume of blood samples being analyzed. The objective of the study was to compare the diagnostic accuracy of a malaria polymerase chain reaction assay, from dried blood spots (DBS, 5 μL) and different volumes of venous blood (50 μL, 200 μL, and 1 mL). The limit of detection of the polymerase chain reaction assay, using calibrated Plasmodium falciparum blood dilutions, showed that venous blood samples (50 μL, 200 μL, 1 mL) combined with Qiagen extraction methods gave a similar threshold of 100 parasites/mL, ∼100-fold lower than 5 μL DBS/Instagene method. On a set of 521 field samples, collected in two different transmission areas in northern Cambodia, no significant difference in the proportion of parasite carriers, regardless of the methods used was found. The 5 μL DBS method missed 27% of the samples detected by the 1 mL venous blood method, but most of the missed parasites carriers were infected by Plasmodium vivax (84%). The remaining missed P. falciparum parasite carriers (N = 3) were only detected in high-transmission areas.     

Detection of Mixed-Species Infections of Plasmodium falciparum and Plasmodium vivax by Nested PCR and Rapid Diagnostic Tests in Southeastern Iran

Coexistence of two species of Plasmodium in a single host has disrupted the diagnosis and treatment of malaria. This study was designed to evaluate the ability of rapid diagnostic test (RDT) kits for the diagnosis of mixed-species malaria infections in southeastern Iran. A total of 100 malaria patients were included in the study out of 164 randomly suspected symptomatic malaria patients from May to November 2012. Nested polymerase chain reaction (PCR) was also used to judge the ability of microscopy versus RDT kits for detecting mixed species. The sensitivity of light microscopy for the detection of mixed-species malaria infections was 16.6% (95% confidence interval [CI] = 3–49.1). Nested PCR revealed 12 patients with mixed-species infection. The CareStart Pv/Pf Combo kit detected 58% of the mixed-species infections, which were determined by nested PCR (sensitivity = 58.3%; 95% CI = 28.5–83.5). For identifying P. falciparum, P. vivax, and mixed-species infections, the concordance rates (kappa statistics) of microscopy and CareStart Pv/Pf Combo kit with nested PCR were 0.76 and 0.79, respectively (P = 0.001). This study underlines the effectiveness of RDT kits to improve the differentiation of mixed-species malaria infections in endemic areas where the prevalence of chloroquine resistance is high.     

Assessing the Performance of CareStart Malaria Pf/Pv Combo Test Against Thick Blood Film in the Diagnosis of Malaria in Northwest Ethiopia

Bivalent rapid diagnostic tests are promising diagnostic tools for Plasmodium falciparum and P. vivax. Their diagnostic performance was evaluated against thick blood smear to assist national malaria control programs. A cross-sectional study was conducted to evaluate the performance of CareStart against thick blood smears among 398 acute febrile patients visiting the Felegeselam Health Center in December of 2011. Thick blood smears were examined under 100× objectives to diagnose Plasmodium species. Similarly, CareStart Malaria Pf/Pv Combo Test was performed as per the manufacturer''s instruction. The ability of CareStart Malaria Pf/Pv Combo Test to diagnose Plasmodium malaria was very good, with 99.8% (95% confidence interval = 97.7–100%) sensitivity and 97.7% (95% confidence interval = 94.6–99.1%) specificity. The sensitivity and specificity of the CareStart Test is comparable with the thick blood smear in diagnosing malaria. Hence, it is preferable to use the CareStart Malaria Pf/Pv Combo Test instead of microscopy in areas where microscopic diagnosis is limited.     

Use of Blood Smears and Dried Blood Spots for Polymerase Chain Reaction–Based Detection and Quantification of Bacterial Infection and Plasmodium falciparum in Severely Ill Febrile African Children

Molecular approaches offer a means of testing archived samples stored as dried blood spots in settings where standard blood cultures are not possible. Peripheral blood films are one suggested source of material, although the sensitivity of this approach has not been well defined. Thin blood smears and dried blood spots from a severe pediatric malaria study were assessed using specific polymerase chain reaction (PCR) primers to detect non-typhoidal Salmonella (NTS; MisL gene), Streptococcus pneumoniae (lytA), and Plasmodium falciparum (18S rRNA). Of 16 cases of NTS and S. pneumoniae confirmed on blood culture, none were positive by PCR using DNA extracts from blood films or dried blood spots. In contrast, four of 36 dried blood spots and two of 178 plasma samples were PCR positive for S. pneumoniae, despite negative bacterial blood cultures, suggesting false positives. Quantitative assessment revealed that the effective concentration of P. falciparum DNA in blood films was three log orders of magnitude lower than for dried blood spots. The P. falciparum kelch13 gene could not be amplified from blood films. These findings question the value of blood PCR-based approaches for detection of NTS and S. pneumoniae, and show that stored blood films are an inefficient method of studying P. falciparum.     

Detection of Plasmodium vivax infection in the Republic of Korea by loop-mediated isothermal amplification (LAMP)

Loop-mediated isothermal amplification (LAMP) is a novel technique that rapidly amplifies target DNA in isothermal conditions. In a previous study, the sensitivities and specificities of LAMP, microscopy, and nested PCR were compared in the context of rapid malaria detection. In the present study, LAMP detected vivax malaria parasites in 115 of 117 microscopically positive samples (sensitivity, 98.3%; 95% CI, 97.4-100%), which agreed well with the nested PCR results (sensitivity, 99.1%; 95% CI: 96.0-100%). No positive cases of malaria were detected by LAMP or nested PCR in 50 consecutive feverish patients other than malaria from malaria endemic areas. LAMP performed on DNA extracted from heat-treated blood had a sensitivity of 93.3% (28/30, 95% CI: 84.4-100%) and specificity of 100% (30/30, 95% CI: 100%). The present study shows that LAMP based assays have high sensitivity, specificity, and amplification efficiencies for Plasmodium vivax detection. The authors recommend that LAMP can be considered as a rapid nucleic acid amplification assay for the molecular diagnosis of P. vivax in both clinical laboratories and malaria clinics in areas where vivax malaria is endemic.     

The burden of pediatric HIV/AIDS in Constanta, Romania: a cross-sectional study

Background

Plasmodium falciparum malaria, is a major health problem in forested tribal belt of central India. Rapid and accurate methods are needed for the diagnosis of P. falciparum. We performed a blinded evaluation of the recently introduced Determine? malaria pf test (Abbott, Laboratories, Japan) compared with microscopy and splenomegaly in children in epidemic prone areas of district Mandla to assess the impact of intervention measures.

Methods

Children aged 2–10 yrs with and without fever were examined for spleen enlargement by medical specialist by establishing a mobile field clinic. From these children thick blood smears were prepared from finger prick and read by a technician. Simultaneously, rapid tests were performed by a field lab attendant. The figures for specificity, sensitivity and predictive values were calculated using microscopy as gold standard.

Results

In all 349 children were examined. The sensitivity and specificity for Determine rapid diagnostic test were 91 and 80% respectively. The positive predictive values (PPV), negative predictive values (NPV) and accuracy of the test were respectively 79, 91 and 85%. On the contrary, the sensitivity and specificity of spleen in detecting malaria infection were 57 and 74 % respectively with PPV of 73%, NPV 59 % and an accuracy of 65%.

Conclusions

Determine? malaria rapid diagnostic test is easier and quicker to perform and has other advantages over microscopy in not requiring prior training of personnel or quality control. Thus, highlighting the usefulness of a rapid antigen test in assessing prevailing malaria situation in remote areas.