Evaluation of Newly Developed Dot-ELISA Test for Identification of Naja-naja sumantrana and Calloselasma rhodostoma Venom Antigens
Snake bite cases in Malaysia most often involve the
species Naja-naja and Calloselasma rhodostoma. In keeping with the
need for a rapid snake venom detection kit in a clinical setting, plate
and dot-ELISA test for the venoms of Naja-naja sumatrana,
Calloselasma rhodostoma and the cobra venom fraction V antigen
was developed. Polyclonal antibodies were raised and further used to
prepare the reagents for the dot-ELISA test kit which was tested in
mice, rabbit and virtual human models. The newly developed dot-
ELISA kit was able to detect a minimum venom concentration of
244ng/ml with cross reactivity of one antibody type. The dot-ELISA
system was sensitive and specific for all three snake venom types in
all tested animal models. The lowest minimum venom concentration
detectable was in the rabbit model, 244ng/ml of the cobra venom
fraction V antigen. The highest minimum venom concentration was
in mice, 1953ng/ml against a multitude of venoms. The developed
dot-ELISA system for the detection of three snake venom types was
successful with a sensitivity of 95.8% and specificity of 97.9%.
[1] Tan NH.Management of Snakebites in Malaysia [Internet].[cited 2010
Sep27].Availablefrom:http://www.tanngethong.com/Management%20of
%20snakebites/management_of_snakebites_in_mala.htm.
[2] Alirol E, Sharma SK,Bawaskar HS,Kuch U,Chappuis F.Snake bite in
South asia: a review.PLos Negl Trop Dis. 2012;4(1):e603.
[3] Kasturiratne A, Wickremasinghe AR, deSilva N,Gunawardena NK,
Pathmeswaran A, et al.The global burden of snakebite:A literature
analysis and modeling based on regional estimates of envenoming and
deaths. PLoS Med 5. 2008;e218.doi:10.1371/journal.pmd.0050218
[4] World Health organization. Rabies and envenomings:A neglected public
health issue.2007. Geneva:WHO.Available at http://www.who.int/blood
products/animal_sera/rabies_envenomings/en/index.html
[5] Gutierrez JM, Theakston RDG, Warell DA. Confronting the neglected
problem of snake bite envenoming: The need for a global partnership.
PLoS Med.2003;3:e150.doi:10.1371/Journal.pmed.0030150.
[6] Warrell DA. Snake bite. Lancet. 2010 Jan 2;375(9708):77-88
[7] Engvall,E and P.perlman. Enzyme linked immunosorbent essay
(ELISA): Quantitation of specific antibodies by enzyme labeled
antiimmunoglobulin in antigen coated tubes. J.Immun.1971,109:129-
135.
[8] Heck FC,Williams JD, Pruett J. Interpretation of spectrophotometric
absorbance values to define results of enzyme -linked immunosorbent
assays. J Clin Microbiol.1980 Apr;11 (4):398-401
[9] Bechi P, Baldini F,Cianchi F. What is the reliability of the portal
spectrophotometer with a fiberoptic probe to detect bilirubin in bile
refluxate? Is sensitivity of Bilitec likely to be improved? Chapter, GER
and gastric motility factors. Available at
http://ww.hon.ch/OESO/books/Vol_5_Eso_Junction/Articles/art118.htm
l
[1] Tan NH.Management of Snakebites in Malaysia [Internet].[cited 2010
Sep27].Availablefrom:http://www.tanngethong.com/Management%20of
%20snakebites/management_of_snakebites_in_mala.htm.
[2] Alirol E, Sharma SK,Bawaskar HS,Kuch U,Chappuis F.Snake bite in
South asia: a review.PLos Negl Trop Dis. 2012;4(1):e603.
[3] Kasturiratne A, Wickremasinghe AR, deSilva N,Gunawardena NK,
Pathmeswaran A, et al.The global burden of snakebite:A literature
analysis and modeling based on regional estimates of envenoming and
deaths. PLoS Med 5. 2008;e218.doi:10.1371/journal.pmd.0050218
[4] World Health organization. Rabies and envenomings:A neglected public
health issue.2007. Geneva:WHO.Available at http://www.who.int/blood
products/animal_sera/rabies_envenomings/en/index.html
[5] Gutierrez JM, Theakston RDG, Warell DA. Confronting the neglected
problem of snake bite envenoming: The need for a global partnership.
PLoS Med.2003;3:e150.doi:10.1371/Journal.pmed.0030150.
[6] Warrell DA. Snake bite. Lancet. 2010 Jan 2;375(9708):77-88
[7] Engvall,E and P.perlman. Enzyme linked immunosorbent essay
(ELISA): Quantitation of specific antibodies by enzyme labeled
antiimmunoglobulin in antigen coated tubes. J.Immun.1971,109:129-
135.
[8] Heck FC,Williams JD, Pruett J. Interpretation of spectrophotometric
absorbance values to define results of enzyme -linked immunosorbent
assays. J Clin Microbiol.1980 Apr;11 (4):398-401
[9] Bechi P, Baldini F,Cianchi F. What is the reliability of the portal
spectrophotometer with a fiberoptic probe to detect bilirubin in bile
refluxate? Is sensitivity of Bilitec likely to be improved? Chapter, GER
and gastric motility factors. Available at
http://ww.hon.ch/OESO/books/Vol_5_Eso_Junction/Articles/art118.htm
l
@article{"International Journal of Medical, Medicine and Health Sciences:53865", author = "A.S. Sikarwar and S. Ambu and T .H. Wong", title = "Evaluation of Newly Developed Dot-ELISA Test for Identification of Naja-naja sumantrana and Calloselasma rhodostoma Venom Antigens", abstract = "Snake bite cases in Malaysia most often involve the
species Naja-naja and Calloselasma rhodostoma. In keeping with the
need for a rapid snake venom detection kit in a clinical setting, plate
and dot-ELISA test for the venoms of Naja-naja sumatrana,
Calloselasma rhodostoma and the cobra venom fraction V antigen
was developed. Polyclonal antibodies were raised and further used to
prepare the reagents for the dot-ELISA test kit which was tested in
mice, rabbit and virtual human models. The newly developed dot-
ELISA kit was able to detect a minimum venom concentration of
244ng/ml with cross reactivity of one antibody type. The dot-ELISA
system was sensitive and specific for all three snake venom types in
all tested animal models. The lowest minimum venom concentration
detectable was in the rabbit model, 244ng/ml of the cobra venom
fraction V antigen. The highest minimum venom concentration was
in mice, 1953ng/ml against a multitude of venoms. The developed
dot-ELISA system for the detection of three snake venom types was
successful with a sensitivity of 95.8% and specificity of 97.9%.", keywords = "ELISA, Venom, SVDK, Naja-naja sumatrana ,
Calloselasma rhodostoma.", volume = "6", number = "8", pages = "376-3", }