Recent advances in the application of molecular biology in filariasis.

Monitoring of filarial parasites in the host and vector has traditionally depended on morphological identification. Recently, species-specific DNA probes have been developed for Brugia malayi, Brugia pahangi and Wuchereria bancrofti. Repeated DNA sequences are useful in developing DNA probes because they evolve more rapidly then coding sequences and their high copy number increases the sensitivity of detection. The Hhal repeated DNA family represents 12% of the total B. malayi DNA. This DNA family is present in species of Brugia (B. malayi, B. timori and B. pahangi) but not W. bancrofti. Sequence analysis of the repeated DNA in B. malayi and B. pahangi has allowed construction of two species-specific DNA probes. These probes were used in a double blind field study in Indonesia. Microfilariae (mf) from infected cats and humans were identified by classical morphological methods and DNA probes. Agreement was found in 98.6% of the 642 samples tested by the two different techniques. Besides mf identification DNA probes can be used to determine the species of infective larvae (L3s) in infected mosquitos. This is useful because the L3s have similar morphology. DNA probes for the identification of W. bancrofti have recently been developed and are in the initial stages of testing in China (Piessens, personal communication) and Egypt (Williams, personal communication). An alternative approach for identification of infected individuals is to detect specific parasite antigens in circulation. A WHO initiative to use either an antigen or antibody assay to replace night blood is presently underway. This approach, if successful would not require the presence of microfilariae, but could detect occult infections.

Evaluation of RNA rich fraction of Litmosoides carinii in induction of protection in infected rats.

The RNA rich fraction of adult L. carinii worms was evaluated in evoking a protective response in infected rats. The RNA immunization was seen to be effective in limiting the microfilaraemia in peripheral blood as well as the adult worm burden. The antibodies to both RNA antigen and adult worm antigen were high in this group of animals at the peak of infection. The RNA immunization was seen to evoke hyperresponsiveness in lymphocytes to mitogens like adult worm antigen, PHA and Con A.

Hybridization between Brugia malayi and Brugia pahangi from south Kalimantan, Indonesia.

Hybridization experiments were carried out between sub-periodic Brugia malayi and B. pahangi recovered from cats originating from the same area in South Kalimantan (Borneo), Indonesia. Five to six week-old immature males and females of both species from Mongolian gerbil (Meriones unguiculatus) intraperitoneal infections were inoculated into the peritoneal cavity of other gerbils. Mating occurred between the species and microfilariae developed into infective third-stage larvae in mosquitoes. Larvae from the mosquitoes developed into adults in the peritoneal cavity of gerbils but microfilariae were not produced since the males were sterile. Back-crossing between hybrids and males and females of the parental species were also done and female hybrids produced microfilariae when crossed with parental males but females from the parental species did not produce microfilariae when mated with the hybrid males. This confirmed that the males were sterile but the females were not. If these species are hybridized in nature and humans are exposed to hybrid third-stage larvae, could this lead to occult filarial infection? These results confirmed previous hybridization experiments of these species orginating from other geographic areas.