How long can stool samples be fixed for an accurate diagnosis of soil-transmitted helminth infection using Mini-FLOTAC?

PLoS Negl Trop Dis. 2015 Apr 7;9(4):e0003698. doi: 10.1371/journal.pntd.0003698. eCollection 2015 Apr.

Abstract

Background: Kato-Katz is a widely used method for the diagnosis of soil-transmitted helminth infection. Fecal samples cannot be preserved, and hence, should be processed on the day of collection and examined under a microscope within 60 min of slide preparation. Mini-FLOTAC is a technique that allows examining fixed fecal samples. We assessed the performance of Mini-FLOTAC using formalin-fixed stool samples compared to Kato-Katz and determined the dynamics of prevalence and intensity estimates of soil-transmitted helminth infection over a 31-day time period.

Methodology: The study was carried out in late 2013 on Pemba Island, Tanzania. Forty-one children were enrolled and stool samples were subjected on the day of collection to a single Kato-Katz thick smear and Mini-FLOTAC examination; 12 aliquots of stool were fixed in 5% formalin and subsequently examined by Mini-FLOTAC up to 31 days after collection.

Principal findings: The combined results from Kato-Katz and Mini-FLOTAC revealed that 100% of children were positive for Trichuris trichiura, 85% for Ascaris lumbricoides, and 54% for hookworm. Kato-Katz and Mini-FLOTAC techniques found similar prevalence estimates for A. lumbricoides (85% versus 76%), T. trichiura (98% versus 100%), and hookworm (42% versus 51%). The mean eggs per gram of stool (EPG) according to Kato-Katz and Mini-FLOTAC was 12,075 and 11,679 for A. lumbricoides, 1,074 and 1,592 for T. trichiura, and 255 and 220 for hookworm, respectively. The mean EPG from day 1 to 31 of fixation was stable for A. lumbricoides and T. trichiura, but gradually declined for hookworm, starting at day 15.

Conclusions/significance: The findings of our study suggest that for a qualitative diagnosis of soil-transmitted helminth infection, stool samples can be fixed in 5% formalin for at least 30 days. However, for an accurate quantitative diagnosis of hookworm, we suggest a limit of 15 days of preservation. Our results have direct implication for integrating soil-transmitted helminthiasis into transmission assessment surveys for lymphatic filariasis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Ancylostomatoidea / isolation & purification
  • Animals
  • Ascaris lumbricoides / isolation & purification
  • Child
  • Feces / parasitology*
  • Female
  • Formaldehyde
  • Helminthiasis / diagnosis*
  • Helminthiasis / epidemiology*
  • Humans
  • Indian Ocean Islands / epidemiology
  • Male
  • Microscopy
  • Prevalence
  • Sensitivity and Specificity
  • Soil / parasitology*
  • Specimen Handling / methods*
  • Tanzania / epidemiology
  • Time Factors
  • Trichuris / isolation & purification

Substances

  • Soil
  • Formaldehyde