FLOTAC: a new sensitive technique for the diagnosis of hookworm infections in humans

Hookworms infect more than 10% of the world's population, but current diagnostic tools have drawbacks. Our objective was to compare the diagnostic performance of three methods (Kato-Katz, ether concentration and FLOTAC techniques) for hookworm diagnosis. Stool samples were obtained from 102 schoolchildren in Côte d'Ivoire. First, a duplicate 41.7 mg Kato-Katz thick smear was prepared. Next, a small portion of stool (mean weight 1.8 g) was preserved in sodium acetate-acetic acid-formalin and forwarded to a European laboratory. These samples were split in three parts, one processed by an ether concentration technique and two by the FLOTAC technique. All samples were examined by experienced technicians for hookworm eggs using light microscopy. The observed hookworm prevalences as assessed by the FLOTAC, Kato-Katz and ether concentration techniques were 65.7%, 51.0% and 28.4%, respectively. Considering the combined results as the diagnostic ‘gold' standard, the FLOTAC technique had a sensitivity of 88.2% compared with 68.4% for the Kato-Katz and 38.2% for the ether concentration techniques. The Kato-Katz method resulted in a significantly higher mean number of eggs per gram of stool (155.8 EPG) compared with the FLOTAC (37.7 EPG) and ether concentration (5.7 EPG) methods. The FLOTAC method shows promise as an important new tool for individual hookworm diagnosis and for rigorous monitoring of helminth control programmes. [Clinical Trial No. ISRCTN21782274

Accuracy of Urine Circulating Cathodic Antigen (CCA) Test for Schistosoma mansoni Diagnosis in Different Settings of Côte d'Ivoire

We aimed to assess the accuracy of a commercially available rapid diagnostic test for the detection of an infection with the blood fluke Schistosoma mansoni in urine. In total, 446 school children from three different settings of south Côte d'Ivoire provided three stool and three urine samples. Stool samples were examined with the widely used Kato-Katz technique and analyzed with a microscope for S. mansoni eggs. Urine samples were examined with a filtration method for S. haematobium eggs and with a rapid diagnostic test for S. mansoni that is based on detecting circulating cathodic antigens (CCA). We used a commercially available test (designated CCA-A) and an experimental formulation (CCA-B). Examination of nine Kato-Katz thick smears per child revealed a prevalence of S. mansoni in the three settings of 32.9%, 53.1%, and 91.8%. The sensitivity of triplicate Kato-Katz from the first stool sample was comparable to a single CCA-A (47.9–94.2% vs. 56.3–89.6%), and significantly higher than the sensitivity of a single CCA-B test (10.4–75.0%). CCA-A showed a considerably lower specificity than CCA-B (76.9–84.2% vs. 96.7–100%). In the settings studied in south Côte d'Ivoire, the CCA-A test holds promise for the diagnosis of S. mansoni, whereas results with CCA-B were suboptimal

Prevalence and risk factors of helminths and intestinal protozoa infections among children from primary schools in western Tajikistan

BACKGROUND: Intestinal parasitic infections represent a public health problem in Tajikistan, but epidemiological evidence is scarce. The present study aimed at assessing the extent of helminths and intestinal protozoa infections among children of 10 schools in four districts of Tajikistan, and to make recommendations for control. METHODS: A cross-sectional survey was carried out in early 2009. All children attending grades 2 and 3 (aged 7-11 years) from 10 randomly selected schools were invited to provide a stool sample and interviewed about sanitary situation and hygiene behaviour. A questionnaire pertaining to demographic and socioeconomic characteristics was addressed to the heads of households. On the spot, stool samples were subjected to duplicate Kato-Katz thick smear examination for helminth diagnosis. Additionally, 1-2 g of stool was fixed in sodium acetate-acetic acid formalin, transferred to a specialized laboratory in Europe and examined for helminths and intestinal protozoa. The results from both methods combined served as diagnostic 'gold' standard. RESULTS: Out of 623 registered children, 602 participated in our survey. The overall prevalence of infection with helminths and pathogenic intestinal protozoa was 32.0% and 47.1%, respectively. There was pronounced spatial heterogeneity. The most common helminth species was Hymenolepis nana (25.8%), whereas the prevalences of Ascaris lumbricoides, hookworm and Enterobius vermicularis were below 5%. The prevalence of pathogenic intestinal protozoa, namely Giardia intestinalis and Entamoeba histolytica/E. dispar was 26.4% and 25.9%, respectively. Almost half of the households draw drinking water from unimproved sources, such as irrigation canals, rivers and unprotected wells. Sanitary facilities were pit latrines, mostly private, and a few shared with neighbours. The use of public tap/standpipe as a source of drinking water emerged as a protective factor for G. intestinalis infection. Protecte spring water reduced the risk of infection with E. histolytica/E. dispar and H. nana. CONCLUSIONS: Our data obtained from the ecological 'lowland' areas in western Tajikistan call for school-based deworming (recommended drugs: albendazole and metronidazole), combined with hygiene promotion and improved sanitation. Further investigations are needed to determine whether H. nana represents a public health problem

Dynamics of Schistosoma haematobium egg output and associated infection parameters following treatment with praziquantel in school-aged children

BACKGROUND: Praziquantel is the drug of choice in preventive chemotherapy targeting schistosomiasis. Increasing large-scale administration of praziquantel requires monitoring of drug efficacy to detect early signs of development of resistance. Standard protocols for drug efficacy monitoring are necessary. Here, we determined the optimal time point for praziquantel efficacy assessment against Schistosoma haematobium and studied the dynamics of infection parameters following treatment. METHODS: Ninety school-aged children from south Cote d'Ivoire with a parasitologically confirmed S. haematobium infection were treated with a single oral dose of praziquantel (40 mg/kg) and followed up for 62 days post-treatment. Urine samples were collected on 23 schooldays during this period and were subjected to visual examination (macrohaematuria), urine filtration and microscopy (S. haematobium eggs) and reagent strip testing (microhaematuria, proteinuria and leukocyturia). RESULTS: Observed cure and egg reduction rates were highly dependent on the time point post-treatment. Egg reduction rates were high (<97%) in weeks 3--9 post-treatment. Cure rates were highest in weeks 6 (92.9%) and 9 (95.0%) post-treatment. The prevalence of infection-associated parameters decreased after treatment, reaching a minimum of 2.4% in weeks 5 (proteinuria) and 7 (leukocyturia) post-treatment, and 16.3% at the end of week 8 (microhaematuria). Macrohaematuria disappeared between weeks 3 and 6 post-treatment. CONCLUSIONS: For monitoring praziquantel efficacy against S. haematobium, we recommend that the cure rate is assessed at week 6 post-treatment. The egg reduction rate can be evaluated earlier, from day 14 post-treatment onwards. Reagent strips are a useful additional tool for evaluating treatment outcomes in areas with high endemicity, preferably at weeks 5 and 6 post-treatment. The delayed decrease of microhaematuria confirms that lesions in the urinary tract persist longer the egg excretion post-treatment

Dynamics of Schistosoma haematobium egg output and associated infection parameters following treatment with praziquantel in school-aged children

BACKGROUND: Praziquantel is the drug of choice in preventive chemotherapy targeting schistosomiasis. Increasing large-scale administration of praziquantel requires monitoring of drug efficacy to detect early signs of development of resistance. Standard protocols for drug efficacy monitoring are necessary. Here, we determined the optimal time point for praziquantel efficacy assessment against Schistosoma haematobium and studied the dynamics of infection parameters following treatment. METHODS: Ninety school-aged children from south Cote d'Ivoire with a parasitologically confirmed S. haematobium infection were treated with a single oral dose of praziquantel (40 mg/kg) and followed up for 62 days post-treatment. Urine samples were collected on 23 schooldays during this period and were subjected to visual examination (macrohaematuria), urine filtration and microscopy (S. haematobium eggs) and reagent strip testing (microhaematuria, proteinuria and leukocyturia). RESULTS: Observed cure and egg reduction rates were highly dependent on the time point post-treatment. Egg reduction rates were high (<97%) in weeks 3--9 post-treatment. Cure rates were highest in weeks 6 (92.9%) and 9 (95.0%) post-treatment. The prevalence of infection-associated parameters decreased after treatment, reaching a minimum of 2.4% in weeks 5 (proteinuria) and 7 (leukocyturia) post-treatment, and 16.3% at the end of week 8 (microhaematuria). Macrohaematuria disappeared between weeks 3 and 6 post-treatment. CONCLUSIONS: For monitoring praziquantel efficacy against S. haematobium, we recommend that the cure rate is assessed at week 6 post-treatment. The egg reduction rate can be evaluated earlier, from day 14 post-treatment onwards. Reagent strips are a useful additional tool for evaluating treatment outcomes in areas with high endemicity, preferably at weeks 5 and 6 post-treatment. The delayed decrease of microhaematuria confirms that lesions in the urinary tract persist longer the egg excretion post-treatment

FLOTAC for the diagnosis of Hymenolepis spp. infection : proof-of-concept and comparing diagnostic accuracy with other methods

Hymenolepis nana is the most common cestode parasitizing humans, yet it is under-diagnosed. We determined the optimal flotation solution (FS) for the diagnosis of this intestinal parasite with the FLOTAC method, and compared its diagnostic accuracy with an ether-concentration technique and the Kato-Katz method. Zinc sulphate (specific gravity 1.20) proved to be the best-performing FS. Using this FS, we detected 65 H. nana infections among 234 fixed fecal samples from Tajik and Sahrawi children (prevalence 27.8 %). The ether-concentration technique detected 40 infections (prevalence 17.1 %) in the same samples. Considering the combined results as a reference, the sensitivities of FLOTAC and ether-concentration were 95.6 % and 58.8 %, respectively. The Kato-Katz method resulted in a prevalence of only 8.7 %. In terms of eggs per gram of stool, a significantly (P >0.05) higher value was obtained with the FLOTAC and Kato-Katz techniques compared to ether-concentration. In another study carried out in China, the FLOTAC method detected six Hymenolepis diminuta infections in 302 fecal samples, whereas five samples were found positive with the Kato-Katz technique. We conclude that FLOTAC is an accurate coprodiagnostic technique for H. nana and H. diminuta, two species which join a growing list of intestinal parasites that can be reliably diagnosed by this techniqu

Comparison of the Flotac-400 Dual Technique and the Formalin-Ether Concentration Technique for Diagnosis of Human Intestinal Protozoon Infection▿

There is a need for accurate diagnosis of intestinal parasite infections in humans, but currently available copromicroscopic techniques have shortcomings, such as low sensitivity. We compared the diagnostic accuracy of a further modified version of the recently developed Flotac technique with that of the widely used formalin-ether concentration technique (FECT) for the detection of intestinal protozoa in human stool samples. Formaldehyde-preserved stool samples from 108 individuals in Côte d'Ivoire were subjected to the Flotac-400 dual technique, using two different flotation solutions (FS), and to the FECT. Stool samples were examined according to computer-generated random lists by an experienced laboratory technician blinded for the results of each method. Both methods detected the same eight intestinal protozoon species. While the Flotac-400 dual technique (results from both FS combined) found higher prevalences of Entamoeba coli (77.8% versus 71.3%, P < 0.001), Blastocystis hominis (20.4% versus 13.0%, P = 0.458), and Giardia intestinalis (8.3% versus 6.5%, P < 0.001), the FECT detected higher prevalences of Entamoeba histolytica/Entamoeba dispar (27.8% versus 20.4%, P = 0.019) and four species of nonpathogenic intestinal protozoa. The diagnostic agreement between the two methods differed considerably depending on the intestinal protozoon investigated (Cohen's kappa measures; range, 0.01 to 0.46). Our study confirmed that the Flotac-400 dual technique can be utilized for the diagnosis of intestinal protozoon infections in humans. Since Flotac is a sensitive technique for the detection of soil-transmitted helminths and Schistosoma mansoni, it might become a viable copromicroscopic technique for the concurrent diagnosis of helminths and intestinal protozoon infections