ABSTRACT
Abstract Background Snakes of the genus Bothrops, popularly known as pit vipers, are responsible for most cases of snakebite in Brazil. Within this genus, Bothrops jararacussu and B. jararaca deserve special attention due to the severity of their bites and for inhabiting densely populated areas. Regarding the treatment of snakebites by Bothrops jararacussu, questions have been raised about the effectiveness of the specific bothropic antivenom in neutralizing myotoxic effects; however, there are no accurate data for humans. Thus, the development of a differential diagnostic kit for this species would be of great interest because it provides, for healthcare professionals, a tool that would allow us to determine whether the accident was caused by B. jararacussu or other species of the genus. It would also make it possible to evaluate the specificity of the treatment and to provide data for epidemiological studies. Methods First, we produced a species-specific polyclonal antibody a potential biomarker of Bothrops jararacussu venom against bothropstoxin-I (BthTx-I), which is also found in smaller quantities in the venoms of B. jararaca from southern Brazil. Results Polyclonal antibodies against bothropstoxin-I could be separated into several species-specific immunoglobulins. Then, aiming to develop a system of safe and standardized immunoassay, we produced monoclonal antibodies. Seven hybridomas were obtained. Five of them were specific to the venom of B. jararacussu and two recognized the venom of B. jararaca from the southeastern population. The use of monoclonal antibodies also made it possible to differentiate B. jararacussu from B. jararaca venom obtained from the southern population. Analyzing the reactivity of monoclonal antibodies against other bothropic venoms, we found mAb Bt-3 to be more specific than others for B. jararacussu venom. Conclusions These results show the potential of BthTx-I for producing monoclonal antibodies that differentiate between B. jararacussu and other Bothrops species venoms.
ABSTRACT
Background Snakes of the genus Bothrops, popularly known as pit vipers, are responsible for most cases of snakebite in Brazil. Within this genus, Bothrops jararacussu and B. jararaca deserve special attention due to the severity of their bites and for inhabiting densely populated areas. Regarding the treatment of snakebites by Bothrops jararacussu, questions have been raised about the effectiveness of the specific bothropic antivenom in neutralizing myotoxic effects; however, there are no accurate data for humans. Thus, the development of a differential diagnostic kit for this species would be of great interest because it provides, for healthcare professionals, a tool that would allow us to determine whether the accident was caused by B. jararacussu or other species of the genus. It would also make it possible to evaluate the specificity of the treatment and to provide data for epidemiological studies. Methods First, we produced a species-specific polyclonal antibody - a potential biomarker of Bothrops jararacussu venom - against bothropstoxin-I (BthTx-I), which is also found in smaller quantities in the venoms of B. jararaca from southern Brazil. Results Polyclonal antibodies against bothropstoxin-I could be separated into several species-specific immunoglobulins. Then, aiming to develop a system of safe and standardized immunoassay, we produced monoclonal antibodies. Seven hybridomas were obtained. Five of them were specific to the venom of B. jararacussu and two recognized the venom of B. jararaca from the southeastern population. The use of monoclonal antibodies also made it possible to differentiate B. jararacussu from B. jararaca venom obtained from the southern population. Analyzing the reactivity of monoclonal antibodies against other bothropic venoms, we found mAb Bt-3 to be more specific than others for B. jararacussu venom. Conclusions These results show the potential of BthTx-I for producing monoclonal antibodies that differentiate between B. jararacussu and other Bothrops species venoms.(AU)
Subject(s)
Animals , Snake Bites , Snakes , Antivenins , Biomarkers , Bothrops , Crotalid Venoms , Antibodies , ImmunoassayABSTRACT
Objective To explore species specificity of 29 Y-SNP loci and to lay the foundation for forensic application.Methods Human DNA and 8 different kinds of common animals' DNA were amplified separately by PCR.The PCR products were analyzed with PAGE.Results Twenty-three out of 29 Y-SNP loci were only amplified by the male human DNA,which indicated a good species specificity.Three loci amplified from human and some animals showed that the length of the product was different between two species.Another three loci amplified from human and animal showed that the length of the fragment was similar between human and animal.Five loci were amplified by PCR products from both male and female samples.Conclusion Most of 29 Y-SNP loci,amplified in male humans,have good species specificity and can be directly used for personal identification and paternity testing.
ABSTRACT
To estimate the postmortem interval (PMI) by using entomological evidence, species identification of forensically important flies is mandatory. However, the traditional species identification method, which relies on the key morphological features of adult flies, is not always available to investigators and has limitations to the immature samples. Because of these limitations, species identification using DNA sequences has long been an issue in the field of forensic entomology. In this review, I have briefly described the basic principles of molecular species identification and phylogenetic analysis and their applications in forensic entomology. I also recommend an experimental and statistical method to identify unknown fly samples obtained from the field.
Subject(s)
Adult , Humans , Base Sequence , Diptera , DNA Barcoding, Taxonomic , Entomology , Forensic Sciences , Korea , Research Personnel , Species SpecificityABSTRACT
Recently forensic scientists have focused on extending the applicability of STR to degraded or tiny amount DNA. Short amplicon may be one of the solutions to these samples and several commercial kits are available for this purpose. Before practical casework, validation is necessary for newly introduced kit. We tried to check how newly introduced STR kit, AmpFLSTR(R) MiniFiler(TM) PCR Amplification Kit, reacts with some animal DNAs. We tried 27 animal DNA samples and checked whether the above kit amplify animal DNA, and next how the band appears on electropherogram. We compared the results with popular STR kits. With AmpFLSTR(R) MiniFiler(TM) PCR Amplification Kit, we could get several bands on electropherogram for some species, but we could not designate proper allelic number compared to allelic ladder for most loci except the following loci, D13S317, D2S1338, D21S11, D18S51, FGA. The D18S51 locus was outstanding in that some species showed definite designated alleles, but the allelic number was not different depending on species. This was the same with another popular STR kit, AmpFLSTR(R) Identifiler(TM) PCR Amplification Kit, which was from the same company. The another kit from another company, PowerPlex(R) 16 System showed different phenomenon with more increased number of amplified bands which were usually differ on size when compared to allelic ladder.
Subject(s)
Animals , Alleles , DNA , Polymerase Chain Reaction , Species SpecificityABSTRACT
Se realizaron prospecciones entomológicas y malacológicas en la Universidad Médica de Camagüey dirigidas a detectar especies de vectores de importancia médico-veterinaria, debido a que en la misma cursan estudios jóvenes procedentes de 40 países, en los cuales de ordinario se reportan importantes enfermedades exóticas para nuestro país. Se identificaron 17 especies con marcada relevancia vectorial, 10 pertenecientes al Phylum Arthropoda Stegomyia aegypti, Anopheles albimanus, Culex quinquefasciatus, Cx. nigripalpus, Ochlerotattus taeniorhynchus y Uranotaenia sapphirina (Familia Culicidae), Musca domestica (Familia Muscidae) y Blatella germánica, Periplaneta americana y Periplaneta brunnea (Familia Blattoptera), así como siete pertenecientes al Phylum Mollusca, cinco dulceacuícolas Depanotrema cimex, Depanotrema sp. (Familia Depanotremidae), Tarebia granifera (Familia Thiaridae), Lymnaea cubensis (Familia Lymnaeidae) y Physa cubensis (Familia Physidae), y dos terrestres Zachrysia auricoma (Familia Camaenidae) y Succinea sp. (Familia Succineinae). Los más abundantes y con mayor variedad de especies resultaron ser los mosquitos. Dichas taxa obliga a diseñar e implementar un adecuado programa de vigilancia y control antivectorial, destinado a evitar la propagación de enfermedades exóticas en nuestro territorio y que están bajo vigilancia epidemiológica.
ABSTRACT
To evaluate species specificity of 20 STRs loci used in our lab. Human DNA and 10 different kinds of common animals' DNA were amplified separately. The PCR products were analysed with PAGE. There are amplified products in DYS388, DYS389, DYS390, D7S809, D13S631 loci of human 'DNA, but not of 10 different kinds of animal DNA tested. There were amplified products in TH01, vWA, AR, CD4, FABP loci for human and animals' DNA, but the lengths of amplified fragment were distinctly different between human and animals. There were products of FES, D12S391, CSF1PO, D19S253, D21S11, FGA, DYS19 loci for human and animals DNA, but the product lengths were not different between human and animals. There were PCR products in SE33 and D11S554 loci for human and animals, their amplified fragment lengths were within the same electrophoretic field. 10 STR loci, such as TH01 etc, were highly specific for human being and the amplified fragments lengths of 8 STR loci, such as FES etc, were different between human and animal tested.