Evaluation of a rapid diagnostic test for assessing the burden of malaria at delivery in India

All pregnant women who came for delivery at a district hospital in Mandla and a civil hospital in Maihar were screened for Plasmodium falciparum (placental parasitemia using a rapid test and microscopy and peripheral and umbilical cord parasitemia using microscopy alone). Two rapid diagnostic tests (RDTs), Paracheck Pf and ParaHITf, were used. At Mandla, the sensitivity and specificity of the Paracheck Pf for P. falciparum were 93% and 84%, respectively. The positive predictive values (PPVs) and negative predictive values (NPVs) were 50% and 99%, respectively. At Maihar, the sensitivity and specificity of the ParaHITf for P. falciparum were 87.5% and 97%, respectively. The PPVs and NPVs were 75.4% and 98.7%, respectively. Placental infection was significantly associated with low birth weight. The RDTs for the identification of P. falciparum were more sensitive in placental blood than the placental blood smear by microscopy. Thus, the RDTs should be useful for rapid assessment of malaria at delivery.     

Use of an HRP2-based rapid diagnostic test to guide treatment of children admitted to hospital in a malaria-endemic area of north-east Tanzania

Objective To compare the performance of the Paracheck? rapid diagnostic test (RDT) with microscopy for diagnosing malaria in hospitalised children. Methods Children aged between 2 months and 13 years with fever were enrolled in the study over 1 year. A standard clinical history and examination were recorded and blood drawn for culture, complete blood count, Paracheck? RDT and double‐read blood slide. Results Of 3639 children enrolled, 2195 (60.3%) were slide positive. The sensitivity and specificity of Paracheck were 97.5% (95% CI 96.9–98.0) and 65.3% (95% CI 63.8–66.9), respectively. There was an inverse relationship between age‐specific prevalence of parasitaemia and Paracheck specificity. In logistic regression model, false‐positive Paracheck results were significantly associated with pre‐admission use of antimalarial drug (OR 1.44, 95% CI 1.16–1.78), absence of current fever (OR 0.64, 95% CI 0.52–0.79) and non‐typhi Salmonella bacteraemia (OR 3.89. 95% CI 2.27–6.63). In spite of high sensitivity, 56/2195 (2.6%) of true infections were Paracheck negative and 8/56 (14.3%) were in patients with >50 000 parasites/μl. Conclusions Paracheck had poor specificity in diagnosing malaria in severely ill children; this was likely to be due to HRP2 persistence following recent parasite clearance. The combination of positive Paracheck and negative blood slide results identified a group of children at high risk of non‐typhi Salmonella infection. While Paracheck was highly sensitive, some high‐density infections were missed. For children with severe febrile illness, at least two reliable negative parasitological test results should be available to justify withholding antimalarial treatment; the optimal choice of these has yet to be identified.     

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.     

Usefulness of a rapid on-site Plasmodium falciparum diagnosis (Paracheck PF) in forest migrants and among the indigenous population at the site of their occupational activities in central India

Logistic, economic, and technical factors limit rapid access to microscopic confirmation of malaria in many tropical countries, including India. The occurrence of high-grade fever and three deaths during the hot summer months in some forest migrants created an emergency situation in Jabalpur in central India. A cheap and rapid malaria test, Paracheck Pf, was tested in this group of migrants in parallel with microscopy. The indigenous population at the site of occupational activities of these migrants approximately 250 km from Jabalpur was also screened by both methods. The results of this field investigation are very encouraging. Among migrants, the test had a sensitivity of 100% and a specificity of 67%. The positive and negative predictive values were 94% and 100%, respectively. Among indigenous population, the corresponding values were 100%, 97.3%, 98.4%, and 100%, respectively, indicating the usefulness of test as a diagnostic tool for providing on-site confirmation of symptomatic diagnosis of Plasmodium falciparum malaria.     

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.     

Challenges in routine implementation and quality control of rapid diagnostic tests for malaria--Rufiji District, Tanzania

Rapid diagnostic tests (RDTs) represent an alternative to microscopy for malaria diagnosis and have shown high sensitivity and specificity in a variety of study settings. Current World Health Organization (WHO) guidelines for quality control of RDTs provide detailed instructions on pre-field testing, but offer little guidance for quality assurance once RDTs are deployed in health facilities. From September 2006 to April 2007, we introduced a histidine-rich protein II (HRP2)-based RDT (Paracheck) for suspected malaria cases five years of age and older in nine health facilities in Rufiji District, Tanzania, to assess sensitivity and specificity of RDTs in routine use at rural health facilities. Thick blood smears were collected for all patients tested with RDTs and stained and read by laboratory personnel in each facility. Thick smears were subsequently reviewed by a reference microscopist to determine RDT sensitivity and specificity. In all nine health facilities, there were significant problems with the quality of staining and microscopy. Sensitivity and specificity of RDTs were difficult to assess given the poor quality of routine blood smear staining. Mean operational sensitivity of RDTs based on reference microscopy was 64.8%, but varied greatly by health facility, range 18.8-85.9%. Sensitivity of RDTs increased with increasing parasite density. Specificity remained high at 87.8% despite relatively poor slide quality. Institution of quality control of RDTs based on poor quality blood smear staining may impede reliable measurement of sensitivity and specificity and undermine confidence in the new diagnostic. There is an urgent need for the development of alternative quality control procedures for rapid diagnostic tests that can be performed at the facility level.     

The diagnosis of Plasmodium falciparum infection in Gambian children, by field staff using the rapid, manual, ParaSight-F test

A rapid immunodiagnostic test for Plasmodium falciparum, the ParaSight-F test, was evaluated in the diagnosis of malaria in 139 children with uncomplicated malaria, who presented at the Medical Research Council's clinic at Basse in Upper River division, The Gambia. The aim was to evaluate the performance and usefulness of the test as a diagnostic method in a malaria-endemic area, when performed by a field worker. Compared with microscopy, the test had a sensitivity of 96.5%, a specificity of 90.5%, a negative predictive value of 94.2% and a positive predictive value of 94.3%. Because of its sensitivity, specificity and simplicity, the ParaSight-F test will be of value in situations where microscopy is not possible.     

Efficacy of a rapid test to diagnose Plasmodium vivax in symptomatic patients of Chiapas, Mexico

OBJECTIVE: To evaluate, under laboratory conditions, the sensitivity and specificity of a rapid diagnostic test (OptiMAL), based on immunoreactive strips, to detect Plasmodium vivax infection in febrile patients in Southern Chiapas, Mexico. MATERIAL AND METHODS: The presence of parasites in blood samples of 893 patients was investigated by Giemsa-stained thick blood smear microscopic examination (gold standard). A blood drop from the same sample was smeared on immunoreactive strips to investigate the presence of the parasite pLDH. Discordant results were resolved by PCR amplification of the parasite's 18S SSU rRNA, to discard infection. RESULTS: OptiMAL had an overall sensitivity of 93.3% and its specificity was 99.5%. Its positive and negative predictive values were 96.5% and 98.9%, respectively. Signal intensity in OptiMAL strips correlated well with the parasitemia density in the blood samples (r = 0.601, p = 0.0001). CONCLUSION: This rapid test had acceptable sensitivity and specificity to detect P. vivax under laboratory conditions and could be useful for malaria diagnosis in field operations in Mexico.     

Evaluation of the performance of CareStart™ Malaria Pf/Pv Combo and Paracheck Pf tests for the diagnosis of malaria in Wondo Genet, southern Ethiopia

Objective

To evaluate the diagnostic performance of CareStart™ Malaria Pf/Pv Combo test relative to microscopy for the diagnosis of falciparum and vivax malaria in Ethiopia.

Methods

668 febrile patients visiting two health centers in Wondo Genet, southern Ethiopia, involved in this study in 2008. Giemsa-stained thin and thick blood smears were prepared and microscopically examined under a 100× oil immersion microscope objective for Plasmodium species identification and determination of parasitaemia, respectively. CareStart™ Malaria Pf/Pv Combo test and Paracheck Pf^® test were performed as per the manufacturers’ instruction.

Findings

The diagnostic validity of CareStart™ Malaria Pf/Pv Combo test for the diagnosis of Plasmodium falciparum were very good with sensitivity of 99.4%, specificity of 98%, positive predictive value of 94.4% and negative predictive value of 99.8%. Sensitivity, specificity, positive predictive value and negative predictive value of the test for the diagnosis of P. vivax were 99.4%, 98.2%, 94.5% and 99.8%, respectively. The diagnostic performance of CareStart™ Malaria Pf/Pv Combo test is comparable to that of Paracheck Pf^® test for the diagnosis of P. falciparum (sensitivity 99.4%, specificity 98.2%).

Conclusion

Although CareStart™ Malaria Pf/Pv Combo test and Paracheck Pf^® test have comparable diagnostic performance for the diagnosis of P. falciparum, CareStart™ Malaria Pf/Pv Combo test has the added advantage of diagnosing P. vivax. Hence, it is preferable to use CareStart™ Malaria Pf/Pv Combo test for the diagnosis of malaria in areas where microscopy is not accessible and where malaria due to P. falciparum and P. vivax are co-endemic as in Ethiopia.     

快速诊断试剂盒在疟疾诊断中的应用效果

目的 探讨疟疾快速诊断试剂盒临床应用效果,为临床应用提供参考。方法 快速诊断试剂盒与镜检法对103 例疟疾、疑似疟疾进行检测对比分析。结果 镜检疟原虫阳性23 例（Pf 11 例、Pv10 例、Pm 和Po 各1 例）,阳性率22.33%;快速诊断试剂盒检测疟原虫阳性22 例（Pf 11 例、Pv 10 例、Pm 1例）,阳性率21.36%,卵形疟未检出。两方法检出阳性率无统计学意义（χ2=0.33,P>0.05）。该试剂的总灵敏性95.65% ,总特异性97.50% ,阳性预测值91.67% ,阴性预测值98.73% ,漏检率4.35% ,误诊率2.50%。结论 疟疾快速诊断试剂盒具有良好的疟疾诊断效果和简便、快速等优点,非常适合镜检技术和条件不足的基层医疗、预防机构开展疟疾检测。     

Comparing the results of light microscopy with the results of PCR method in the diagnosis of Plasmodium vivax

BACKGROUND AND OBJECTIVES: Although polymerase chain reaction (PCR) is a new technique in the diagnosis of malaria with very high accuracy; light microscopy is still conventional diagnostic method in Iran. In this study we checked the accuracy of light microscopy using the results of PCR as gold standard in Iran. METHODS: The blood samples were collected from 124 febrile cases in Kahnooj district. The blood slides were read by microscopists, and double checked by experts in provincial referral laboratory. DNA samples were processed by PCR to amplify species-specific sequences of 18s subunit ribosomal ribonucleic acid (18ssrRNA) genes of Plasmodium vivax and P. falciparum. RESULTS: The sensitivity and specificity of microscopy in the detection of Plasmodium spp infection were 77% (95% CI: 46-94%) and 100% (95% CI: 95-100%), correspondingly. Also, the estimated positive and negative predictive values were 100% (95% CI: 66-100%) and 97% (95% CI: 91-99%), respectively. INTERPRETATION AND CONCLUSION: According to these results, we believe that the accuracy of light microscopy in the diagnosis of malaria in Kahnooj was acceptable. Expert micorscopists in endemic areas of Iran such as Kahnooj and available equipments in one hand and expensive PCR test on the other hand may convince that in current situation we do not have to change the diagnostic method.     

Automated detection of malaria pigment: feasibility for malaria diagnosing in an area with seasonal malaria in northern Namibia

OBJECTIVE: To evaluate the feasibility of automated malaria detection with the Cell-Dyn 3700 (Abbott Diagnostics, Santa Clara, CA, USA) haematology analyser for diagnosing malaria in northern Namibia. METHODS: From April to June 2003, all patients with a positive blood smear result and a subset of patients with no suspicion of malaria were included. Blood smear and a venous blood sample (to determine haemoglobin, platelet and malaria pigment levels) were collected from each patient. Malaria pigment test characteristics, correlations with blood parameters and pigment clearance time were calculated. Finally, a subset of blood samples was run twice to evaluate the consistency of test outcome. RESULTS: Two hundred and eight patients were included. Ninety had a positive blood smear result of which 84 tested positive for malaria pigment and 118 patients had a negative blood smear result of which four tested positive for malaria pigment. Test characteristics as compared with microscopy were as follows: sensitivity 0.93, specificity 0.97, positive predictive value 0.95, negative predictive value 0.95. Rerun of the blood samples resulted in a change of diagnosis in 14%. After 4 weeks, 33% of patients with an initially positive pigment result still tested positive. Malaria pigment was found to be negatively correlated with haemoglobin. CONCLUSIONS: Automated detection of malaria pigment is a useful diagnostic tool in this semi-rural area. In low-risk malaria season, the test can be used for diagnosing malaria because of the high sensitivity. In high-risk malaria season, the test can be used for excluding malaria in case of a negative pigment result because of the high specificity.     

Evaluation of a rapid diagnostic test, 'Determine malaria pf', in epidemic-prone, forest villages of central India (Madhya Pradesh)

A rapid, immunochromatographic test for malaria diagnosis, 'Determine malaria pf', was evaluated by a field team in the epidemic-affected, forest setting of Chhindwara district, in Madhya Pradesh, central India. In all, 526 fever cases were screened for Plasmodium falciparum in October or November, 1999. Those found to be infected were treated with sulfadoxine-pyrimethamine and primaquine. Using microscopy as the gold standard, the new test had a sensitivity of 98% and a specificity of 87%. The positive and negative predictive values were 88% and 98%, respectively. Although follow-up of 64 subjects on day 7 post-treatment revealed that 20% of those who then appeared smear-negative were still antigenaemic, 34% of the subjects were still smear-positive, for asexual parasites, at that time. The Determine test was found to be very easy to perform and the results could be read reliably by field workers, without any supervision. The ease of use of the test indicates that it could be useful in the management of malaria, particularly in remote and inaccessible areas, provided that its accuracy can be assured and that it can be made affordable.     

Field studies on the sensitivity and specificity of an immunochromatographic test for the detection of Plasmodium falciparum malaria in tribal areas of Orissa

A rapid immunodiagnostic test developed by an Australian Biotechnology company for the diagnosis of Plasmodium falciparum in the peripheral blood has been evaluated in the field for its sensitivity, specificity and efficacy in comparison to microscopic examination. The results showed that the tests sensitivity, specificity and efficacy were 98.2, 96.9 and 97.5 per cent respectively. The positive and negative predictive values of the test were 96.4 and 98.4 per cent respectively. The test when compared to the conventional microscopy did not show any statistically significant difference suggesting that the two diagnostic methods are equally good. The test performed did not show cross-reactions with other parasite species. It is a simple and rapid field diagnostic method, which does not require any expensive laboratory equipment or skilled personnel.     

Quality Assurance of Rapid Diagnostic Tests for Malaria in Routine Patient Care in Rural Tanzania

Histidine-rich protein II (HRP2)-based malaria rapid diagnostic tests (RDTs) have shown high sensitivity and specificity for detecting Plasmodium falciparum malaria in a variety of study settings. However, RDTs are susceptible to heat and humidity and variation in individual performance, which may affect their use in field settings. We evaluated sensitivity and specificity of RDTs during routine use for malaria case management in peripheral health facilities. From December 2007 to October 2008, HRP2-based ParaHIT-f RDTs were introduced in 12 facilities without available microscopy in Rufiji District, Tanzania. Health workers received a single day of instruction on how to perform an RDT and thick blood smear. Job aids, Integrated Management of Childhood Illness guidelines, and national malaria treatment algorithms were reviewed. For quality assurance (QA), thick blood smears for reference microscopy were collected for 2 to 3 days per week from patients receiving RDTs; microscopy was not routinely performed at the health facilities. Slides were stained and read centrally within 72 hours of collection by a reference microscopist. When RDT and blood smear results were discordant, blood smears were read by additional reference microscopists blinded to earlier results. Facilities were supervised monthly by the district laboratory supervisor or a member of the study team. Ten thousand six hundred fifty (10,650) patients were tested with RDTs, and 51.5% (5,488/10,650) had a positive test result. Blood smear results were available for 3,914 patients, of whom 40.1% (1,577/3,914) were positive for P. falciparum malaria. Overall RDT sensitivity was 90.7% (range by facility 85.7–96.5%) and specificity was 73.5% (range 50.0–84.3%). Sensitivity increased with increasing parasite density. Successful implementation of RDTs was achieved in peripheral health facilities with adequate training and supervision. Quality assurance is essential to the adequate performance of any laboratory test. Centralized staining and reading of blood smears provided useful monitoring of RDT performance. However, this level of QA may not be sustainable nationwide.     

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 malaria rapid diagnostic tests for the diagnosis of P. falciparum and non-P. falciparum infections

The objective of this study was to evaluate various malaria rapid diagnostic tests as a tool in the detection of P. falciparum and non-P. falciparum infections in field conditions. Four field surveys were conducted in malaria-endemic areas of Palawan and Davao del Norte, Philippines to validate the various rapid diagnostic tests, namely Diamed OptiMAL 48 (DiaMed AG, Switzerland), ParaHIT f (Span Diagnostics, India), Orchid OptiMAL, and Paracheck Pf (both from Orchid Biomedical Systems, India). The results of the various rapid diagnostic tests were compared to those of microscopy. Sensitivity, specificity and detection rates according to the level of parasitemia were used as parameters to describe the performance of the various rapid diagnostic tests in the field. Practical and operational assessments were also done. The results of the study show that the sensitivity and detection rates were generally lower than previously reported, with sensitivities ranging from 4.8% to 20.6%, except for Diamed OptiMAL 48, which had sensitivities of 78.8% to 96.8%, and detection rates of 50.0% to 96.8%. The rest had detection rates ranging from 0.0% to 50.0%. All the specificities ranged from 18.2% to 100.0%. Improper conditions at the time of manufacturing, storage, transport, and utilization may affect the validity of the results. Rapid diagnostic tests for malaria provide practical means of detecting malarial infections, especially in endemic areas. However, issues regarding variability in performance must to be addressed before they can be used as mainstream diagnostic tools.     

镜检、抗原检测和核酸检测方法在疟疾诊断中的应用比较

目的 比较镜检、抗原检测(RDT)和核酸检测(PCR)三种方法对疟原虫的检测效果,为基层选择合适的 诊断方法提供依据。 方法 收集腾冲市 2015-2018 年发热病人的血样进行疟疾检测,以确诊结果为标准,对比分析镜检、RDT 和 PCR 三种疟疾检测方法的敏感性、特异性、阳性预测值、阴性预测值等指标。 结果 610 份血样中,阴性 295 份,阳性 315 份,其中恶性疟 67 份、间日疟 245 份、混合感染 2 份、三日疟 1 份。 与确诊结果比较,镜检、RDT 和 PCR 的灵敏度分别为 95. 87%、94. 60%和 99. 37%,特异度均为 100%;假阴性率分别为 4. 13%、5. 40%和 0. 63%,阴性预 测值分别为 95. 78%、94. 55%和 99. 33%;假阳性率均为 0,阳性预测值均为 100%;三种方法与确诊结果的总符合率分别 为 97. 87%、97. 70%和 99. 67%,Kappa 检验结果显示均与确诊结果高度一致(P 均<0. 001);对单一虫种恶性疟的检测, 镜检、RDT 和 PCR 的符合率分别为 88. 06%、100%和 97. 01%;对其他三种疟原虫检测的符合率,分别为 97. 97%、 93. 9%和 100%。 结论 三种检测方法均具有较高的敏感性和特异性,但综合考虑当前防治工作实际,抗原检测(RDT) 更适宜在基层推广和使用。     

Rapid diagnostic tests for malaria at sites of varying transmission intensity in Uganda

BACKGROUND: In Africa, fever is often treated presumptively as malaria, resulting in misdiagnosis and the overuse of antimalarial drugs. Rapid diagnostic tests (RDTs) for malaria may allow improved fever management. METHODS: We compared RDTs based on histidine-rich protein 2 (HRP2) and RDTs based on Plasmodium lactate dehydrogenase (pLDH) with expert microscopy and PCR-corrected microscopy for 7000 patients at sites of varying malaria transmission intensity across Uganda. RESULTS: When all sites were considered, the sensitivity of the HRP2-based test was 97% when compared with microscopy and 98% when corrected by PCR; the sensitivity of the pLDH-based test was 88% when compared with microscopy and 77% when corrected by PCR. The specificity of the HRP2-based test was 71% when compared with microscopy and 88% when corrected by PCR; the specificity of the pLDH-based test was 92% when compared with microscopy and >98% when corrected by PCR. Based on Plasmodium falciparum PCR-corrected microscopy, the positive predictive value (PPV) of the HRP2-based test was high (93%) at all but the site with the lowest transmission rate; the pLDH-based test and expert microscopy offered excellent PPVs (98%) for all sites. The negative predictive value (NPV) of the HRP2-based test was consistently high (>97%); in contrast, the NPV for the pLDH-based test dropped significantly (from 98% to 66%) as transmission intensity increased, and the NPV for expert microscopy decreased significantly (99% to 54%) because of increasing failure to detect subpatent parasitemia. CONCLUSIONS: Based on the high PPV and NPV, HRP2-based RDTs are likely to be the best diagnostic choice for areas with medium-to-high malaria transmission rates in Africa.     

Evaluation of direct agglutination test (DAT) as an immunodiagnostic tool for diagnosis of visceral leishmaniasis in Nepal

Before field application of the direct agglutination test (DAT) for leishmaniasis, it was assessed as a diagnostic tool. Fifteen confirmed visceral leishmaniasis cases (bone marrow aspiration positive), 120 tuberculosis, 58 leprosy, 15 malaria, 26 intestinal parasitic infection cases, 24 endemic healthy controls from adjacent to the study area, and 18 controls from Kathmandu (who had never visited the VL endemic areas) were tested for anti-leishmanial antibody agglutination titers. Two of the tuberculosis cases were positive for anti-leishmanial agglutinating antibodies at 1:800. All the visceral leishmaniasis confirmed cases were reactive to anti-leishmanial antibody at > or = 1:3,200. Other specimens were negative for serology. The sensitivity of the direct agglutination test was 100% and the specificity was 99.2%. The direct agglutination test had positive and negative predictive values of 100% and 99.2% respectively. The direct agglutination test has been found to be simple, rapid, reliable, economic, safe and adaptable to micro-techniques using microtiter plates. It is specific and sensitive. The direct agglutination test is simple enough for it to be performed in a field laboratory.