Quantitation of Indian krait (Bungarus caeruleus) venom in human specimens of forensic origin by indirect competitive inhibition enzyme-linked immunosorbent assay

J AOAC Int. 2006 Sep-Oct;89(5):1360-6.

Abstract

An indirect competitive inhibition enzyme-linked immunosorbent assay was reported to detect krait venom in human specimens of forensic origin. Polyclonal anti-krait venom antibodies were characterized by indirect antibody capture assay. The calibration plot was constructed based on linear regression analysis (y = 72.85 - 12.29x, r(2) = 0.98) with concentration ranges from 0.013 to 1000 ng/well of krait venom with a limit of detection of 0.2 ng/mL in the assay system. The IC50 (inhibitory concentration at 50% displacement) value of krait venom was observed to be 70 ng. Spiking studies indicated recoveries of 95-100% and 94-100% when various concentrations of krait venom were spiked to rat tissues (skin, liver, and kidneys) and pooled human serum, respectively. Polyclonal anti-krait venom antibodies showed no cross-reactivity with cobra and viper venom when tested in the assay system. The coefficient of variation of various concentrations of working range in intra-assay (n = 6) was <5%, whereas in interassay (n = 6) it was observed to be < or 7%. Further, the method was used to quantitate krait venom in human autopsy and biopsy specimens of forensic origin. Concentration of krait venom was found to be in the range of 4-172 ng/100 mg skin or skin scrapings and 64-378 ng/mL blood or serum. The methodology may find application in forensic laboratories to assess the cause of death in the cases of krait-bite victims.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adolescent
  • Adult
  • Animals
  • Antibodies / isolation & purification
  • Autopsy
  • Biopsy
  • Bungarus*
  • Child
  • Elapid Venoms / analysis*
  • Elapid Venoms / immunology
  • Enzyme-Linked Immunosorbent Assay / methods*
  • Enzyme-Linked Immunosorbent Assay / standards
  • Enzyme-Linked Immunosorbent Assay / statistics & numerical data
  • Female
  • Forensic Medicine / methods
  • Humans
  • Immunization
  • India
  • Male
  • Middle Aged
  • Rabbits
  • Sensitivity and Specificity
  • Snake Bites / diagnosis

Substances

  • Antibodies
  • Elapid Venoms