A single injection of 19 kDa carboxy-terminal fragment of Plasmodium yoelii merozoite surface protein 1 (PyMSP119) formulated with Montanide ISA and CpG ODN induces protective immune response in mice.

Objective: To investigate the efficacy of a vaccine formulation of the 19 kDa conserved carboxyl-terminal fragment of Plasmodium yoelii merozoite surface protein-1 (PyMSP119) formulated with CpG ODN 1826 and Montanide ISA51 or ISA720 when used to immunize mice by a single injection. Methods: Groups of BALB/c mice were immunized parenterally with one, two or four injections with PBS or PyMSP119 formulated with CpG ODN in ISA51 or ISA720. Sera were collected weekly and assessed for total IgG and IgG subclass titers. Protection was tested by challenge infection with P. yoelii YM. Results: Interestingly, single injection immunization showed the same kinetics of antibody responses as two- or four-injection immunization. However, the peak antibody response induced by PyMSP119 in CpG ODN and ISA51 appeared earlier than that induced by PyMSP119 in CpG ODN and ISA720 (28 days vs 41 days). At day 63 after the first injection, the PyMSP119-specific IgG antibody levels by single injection and four- injection immunizations were not different. However, the levels of the IgG2a antibody subclass were significantly lower by single injection immunization with PyMSP119 in CpG ODN and ISA720. The antibodies were sustained at high levels for at least 20 weeks. After challenge infection, all mice immunized by a single injection of PyMSP119 in CpG ODN and ISA51 survived with low-grade parasitemia, while 50% of mice immunized with PyMSP119 in CpG ODN and ISA720 died with high levels of parasitemia. Conclusion: These findings suggest that MSP119 immunization by a single injection can induce protective immunity, particularly when formulated with an appropriate strong adjuvant.

Dengue hemorrhagic fever: knowledge, attitude and practice in Ang Thong Province, Thailand.

A cross-sectional study was carried out between July 1998 and June 1999 to identify dengue virus-infected patients under age 15 admitted to seven government hospitals in Ang Thong Province, a central region of Thailand, and to assess the knowledge, attitude, and practice (KAP) of their care takers. To differentiate dengue cases, clinical evaluation and laboratory diagnosis were used. Serum samples were collected from 90 admitted children and also from 80 healthy students. The dengue cases were classified as dengue fever (9 cases, 12.2%) and dengue hemorrhagic fever (DHF: 65 cases, 87.8%). Nine patients had dengue shock syndrome, but no death occurred. With serological confirmation, primary antibody response was observed in 8 (11.3%) and definite secondary infection in 49 (69%). Out of 41 serum samples, 14 (34.1%) were positive for dengue virus isolation: dengue serotypes 1, 2 or 3. A total of 131 care takers of enrolled children were interviewed in the context of KAP in DHF. The majority of them were mothers with primary school education level. Half of the care takers were workers. DHF knowledge of the care takers of the dengue cases, non-cases, and healthy students was almost the same. However, the care takers of dengue cases recognized petechiae as a danger sign, p-value of 0.006. They had a higher response in prevention, control and treatment of DHF than the other two groups after their children were admitted to hospital, p-value of 0.000. The results indicated that DHF remains a public health problem in this area and the people need more understanding of the disease. Continuous campaigns are required for community participation so as to prevent and control DHF successfully.

An improved method for concentrating rotavirus from water samples

A modified adsorption-elution method for the concentration of seeded rotavirus from water samples was used to determine various factors which affected the virus recovery. An enzyme-linked immunosorbent assay was used to detect the rotavirus antigen after concentration. Of the various eluents compared, 0.05M glycine, pH 11.5 gave the highest rotavirus antigen recovery using negatively charged membrane filtration whereas 2.9 percent tryptose phosphate broth containing 6 percent glycine; pH 9.0 was found to give the greatest elution efficiency when a positively charged membrane was used. Reconcentration of water samples by a speedVac concentrator showed significantly higher rotavirus recovery than polyethylene glycol precipitation through both negatively and positively charged filters (p-value <0.001). In addition, speedVac concentration using negatively charged filtration resulted in greater rotavirus recovery than that using positively charged filtration (p-value = 0.004). Thirty eight environmental water samples were collected from river, domestic sewage, canals receiving raw sewage drains, and tap water collected in containers for domestic use, all from congested areas of Bangkok. In addition, several samples of commercial drinking water were analyzed. All samples were concentrated and examined for rotavirus antigen. Coliforms and fecal coliforms (0->1,800 MPN/100 ml) were observed but rotavirus was not detected in any sample. This study suggests that the speedVac reconcentration method gives the most efficient rotavirus recovery from water samples