Quantitation of Indian krait (Bungarus caeruleus) venom in human specimens of forensic origin by indirect competitive inhibition enzyme-linked immunosorbent assay.
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.
Brunda
Ganneru
G
Osmania University, University College of Science, Department of Biochemistry, Hyderabad 7, India.
Rao
Beedu Sashidhar
BS
Sarin
Rajendra Kumar
RK
D000293:Adolescent; D000328:Adult; D000818:Animals; D000906:Antibodies / Q000302:isolation & purification; D001344:Autopsy; D001706:Biopsy; D017838:Bungarus; D002648:Child; D004546:Elapid Venoms / Q000032:analysis* / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods* / Q000592:standards / Q000706:statistics & numerical data; D005260:Female; D005554:Forensic Medicine / Q000379:methods; D006801:Humans; D007114:Immunization; D007194:India; D008297:Male; D008875:Middle Aged; D011817:Rabbits; D012680:Sensitivity and Specificity; D012909:Snake Bites / Q000175:diagnosis
D000906:Antibodies; D004546:Elapid Venoms