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ObjectivesThis study aims to develop a high-sensitivity antibody diagnostic kit that will enable a rapid and accurate detection of Cryptospofidium parvum and Giardia lamblia in patients with diarrhea.MethodsThe cultivated C. parvum oocysts and G. lamblia cysts in each calf and dog were injected to mice to obtain antibodies, which were titrated. Spleen cells of the immunized mouse were separated and blended with myelomas to produce hybrid cell lines that form monoclonal antibodies. Using ELISA method, antibodies that specifically respond to C. parvum and G.lamblia were then selected. The cells were injected into the abdominal cavity of a BALB/c mouse to isolate hydrops abdominis containing high level of antibodies. The IgG antibody was purified using protein G gel.ResultsThe detection limit of monoclonal antibodies for Cryptosporidium parvum and Giardia lamblia was 125 oocysts/mL and 1250 cysts/mL, respectively. In addition, during testing they did not show cross-reactivity to viruses (n = 15), bacteria (n =17), and parasites (n = 9).ConclusionThe rapid diagnostic antibody kit developed in this study, which specifically responds to C. parvum and G. lamblia, will be useful in detecting and monitoring diarrheal infections.

Background It is almost an axiom that in the African highlands (above 1,500 m) transmission of Plasmodium falciparum is limited primarily by low ambient temperature and that small changes in temperature could result in temporary favorable conditions for unstable transmission within populations that have acquired little functional immunity. The pattern of malaria transmission in the highland plateau ecosystems is less distinct due to the flat topography and diffuse hydrology resulting from numerous streams. The non-homogeneous distribution of larval breeding habitats in east African highlands obviously affects Anopheles spatial distribution which, consequently, leads to heterogeneous human exposure to malaria. Another delicate parameter in the fragile transmission risk of malaria in the highlands is the rapid loss of primary forest due to subsistence agriculture. The implication of this change in land cover on malaria transmission is that deforestation can lead to changes in microclimate of both adult and larval habitats hence increase larvae survival, population density, and gametocytes development in adult mosquitoes. Deforestation has been documented to enhancing vectorial capacity of Anopheles gambiae by nearly 100% compared to forested areas. Method The study was conducted in five different ecosystems in the western Kenya highlands, two U-shaped valleys (Iguhu, Emutete), two V-shaped valleys (Marani, Fort Ternan), and one plateau (Shikondi) for 16 months among 6- to 15-year-old children. Exposure to malaria was tested using circumsporozoite protein (CSP) and merozoite surface protein immunochromatographic antibody tests. Malaria parasite was examined using different tools, which include microscopy based on blood smears, rapid diagnostic test based on HRP 2 proteins, and serology based on human immune response to parasite and vector antigens have been also examined in the highlands in comparison with different topographical systems of western Kenya. Results The results suggested that changes in the topography had implication on transmission in highlands of western Kenya and appropriate diagnosis, treatment, and control tool needed to be considered accordingly. Both plateau and U-shaped valley found to have higher parasite density than V-shaped valley. People in V-valley were less immune than in plateau and U-valley residents. Conclusion Topography diversity in western Kenya highlands has a significant impact on exposure rates of human to malaria vectors and parasite. The residents of V-shaped valleys are at risk of having explosive malaria outbreaks during hyper-transmission periods due to low exposure to malaria parasite; hence, they have low immune response to malaria, while the U-shaped valleys have stable malaria transmission, therefore, the human population has developed immunity to malaria due to continuous exposure to malaria.

Background Diagnosing Plasmodium falciparum malaria during pregnancy is a great challenge for clinicians because of the low density of parasites in the peripheral blood and parasite sequestration in the placenta. Nevertheless, few data on the use of malaria rapid diagnostic test (RDT) during pregnancy have been published. Methods P. falciparum infections were assessed in 156 febrile pregnant women by microscopic examination of their blood smears and by RDT and polymerase chain reactions (PCR). In addition, 150 women were assessed at the time of delivery by microscopy, RDT, PCR and placental histology investigations. The study was conducted at the Gadarif Hospital, Eastern Sudan. The SD Bioline P. f / P. v (Bio Standard Diagnostics, Gurgaon, Korea) RDT kit was evaluated in this study. Results Among the febrile pregnant women, 17 (11.0%), 26 (16.7%) and 18 (11.5%) positive cases of P. falciparum were detected by microscopy, RDT, and PCR, respectively. The sensitivity and specificity of the microscopy was 94.4% and 100%, respectively. The corresponding values for RDT evaluation were 83.3% and 92.0%, as compared with PCR as the gold standard. While there were no detected cases of malaria by microscopic examination of blood smears, 27 (18.0%), 21(14.0%) and 46 (30.7%) out of the 150 placentae investigated had P. falciparum as determined by RDT, PCR, and histology, respectively. The sensitivity and specificity for RDT was 17.4% and 81.7%, respectively. The corresponding values for PCR were 6.5% and 82.7%, where histology was used as the gold standard. Conclusions The RDT kit used in this study has poor performance for peripheral and placental P. falciparum malaria detection in this setting. Virtual slides The virtual slide(s) for this article can be found here: http://www.diagnosticpathology.diagnomx.eu/vs/1092363465928479