Parasitological diagnosis of onchocerciasis: comparisons of incubation media and incubation times for skin snips.

Tissue culture fluid NCTC 135 (Hank's base) was compared to water and to saline as incubation media for the detection of microfilariae of Onchocerca volvulus in skin snips. NCTC 135 allowed detection of significantly more positive persons than did water (P less than 0.001) or saline (P less than 0.05) when two snips per person were incubated for periods of 0.5 or 24 hours. In addition, snips containing microfilariae were incubated in NCTC 135 or in saline and the number of emerged microfilariae was determined at various intervals of time up to 24 hours. After incubation, snips were either fixed in 10% formalin, serially sectioned, and the microfilariae counted, or they were digested in collagenase solution to free unemerged microfilaire. Of the total number of microfilariae present in the snips, 43.9% +/- 18.5, 80.2% +/- 22.2, 83.0% +/- 19.5, and 85.3% +/- 18.0 had emerged by 0.5, 4, 8, and 24 hours of incubation, respectively. Of the microfilariae that remained in the skin after incubation, most were located deep in the dermis.