Hallucinogenic mushroom is a kind of toxic strain containing psychoactive tryptamine substances such as psilocybin,psilocin and ibotenic acid,etc.The mushrooms containing hallucinogenic components are various,widely d...Hallucinogenic mushroom is a kind of toxic strain containing psychoactive tryptamine substances such as psilocybin,psilocin and ibotenic acid,etc.The mushrooms containing hallucinogenic components are various,widely distributed and lack of standard to define,which made a great challenge to identification.Traditional identification methods,such as morphology and toxicology analysis,showed shortcomings in old or processed samples,while the DNA-based identification of hallucinogenic mushrooms would allow to identify these samples due to the stability of DNA.In this paper,four primer sets are designed to target Psilocybe cubensis DNA for increasing resolution of present identification method,and the target markers include largest subunit of RNA polymerase II(marked as PC-R1),psilocybin-related phosphotransferase gene(marked as PC-PT),glyceraldehyde 3-phosphate dehydrogenase(marked as PC-3)and translation EF1α(marked as PC-EF).Real-time PCR with high-resolution melting(HRM)assay were used for the differentiation of the fragments amplified by these primer sets,which were tested for specificity,reproducibility,sensitivity,mixture analysis and multiplex PCR.It was shown that the melting temperatures of PC-R1,PC-PT,PC-3 and PC-EF of P.cubensis were(87.93±0.12)℃,(82.21±0.14)℃,(79.72±0.12)℃ and(80.11±0.19)℃ in our kinds of independent experiments.Significant HRM characteristic can be shown with a low concentration of 62.5pg/µL DNA sample,and P.cubensis could be detected in mixtures with Homo sapiens or Cannabis sativa.In summary,the method of HRM analysis can quickly and specifically distinguish P.cubensis from other species,which could be utilized for forensic science,medical diagnosis and drug trafficking cases.展开更多
Mushroom poisoning is the main cause of mortality in food poisoning incidents in China.Although some responsible mushroom species have been identified,some were identified inaccuratly.This study investigated and analy...Mushroom poisoning is the main cause of mortality in food poisoning incidents in China.Although some responsible mushroom species have been identified,some were identified inaccuratly.This study investigated and analyzed 102 mushroom poisoning cases in southern China from 1994 to 2012,which involved 852 patients and 183 deaths,with an overall mortality of 21.48%.The results showed that 85.3%of poisoning cases occurred from June to September,and involved 16 species of poisonous mushroom:Amanita species(A.fuliginea,A.exitialis,A.subjunquillea var.alba,A.cf.pseudoporphyria,A.kotohiraensis,A.neoovoidea,A.gymnopus),Galerina sulciceps,Psilocybe samuiensis,Russula subnigricans,R.senecis,R.japonica,Chlorophyllum molybdites,Paxillus involutus,Leucocoprinus cepaestipes and Pulveroboletus ravenelii.Six species(A.subjunquillea var.alba,A.cf.pseudoporphyria,A.gymnopus,R.japonica,Psilocybe samuiensis and Paxillus involutus)are reported for the first time in poisoning reports from China.Psilocybe samuiensis is a newly recorded species in China.The genus Amanita was responsible for 70.49%of fatalities;the main lethal species were A.fuliginea and A.exitialis.Russula subnigricans caused 24.59%of fatalities,and five species showed mortality>20%(A.fuliginea,A.exitialis,A.subjunquillea var.alba,R.subnigricans and Paxillus involutus).Mushroom poisoning symptoms were classified from among the reported clinical symptoms.Seven types of mushroom poisoning symptoms were identified for clinical diagnosis and treatment in China,including gastroenteritis,acute liver failure,acute renal failure,psychoneurological disorder,hemolysis,rhabdomyolysis and photosensitive dermatitis.展开更多
基金supported by grants from the Shanghai Areas of Development for Society Planning Projects[grant number 19dz1200600]the National Natural Science Foundation of China[grant numbers 81930056 and 81625013]the National Youth Talent Support Program[grant number WRQB2019].
文摘Hallucinogenic mushroom is a kind of toxic strain containing psychoactive tryptamine substances such as psilocybin,psilocin and ibotenic acid,etc.The mushrooms containing hallucinogenic components are various,widely distributed and lack of standard to define,which made a great challenge to identification.Traditional identification methods,such as morphology and toxicology analysis,showed shortcomings in old or processed samples,while the DNA-based identification of hallucinogenic mushrooms would allow to identify these samples due to the stability of DNA.In this paper,four primer sets are designed to target Psilocybe cubensis DNA for increasing resolution of present identification method,and the target markers include largest subunit of RNA polymerase II(marked as PC-R1),psilocybin-related phosphotransferase gene(marked as PC-PT),glyceraldehyde 3-phosphate dehydrogenase(marked as PC-3)and translation EF1α(marked as PC-EF).Real-time PCR with high-resolution melting(HRM)assay were used for the differentiation of the fragments amplified by these primer sets,which were tested for specificity,reproducibility,sensitivity,mixture analysis and multiplex PCR.It was shown that the melting temperatures of PC-R1,PC-PT,PC-3 and PC-EF of P.cubensis were(87.93±0.12)℃,(82.21±0.14)℃,(79.72±0.12)℃ and(80.11±0.19)℃ in our kinds of independent experiments.Significant HRM characteristic can be shown with a low concentration of 62.5pg/µL DNA sample,and P.cubensis could be detected in mixtures with Homo sapiens or Cannabis sativa.In summary,the method of HRM analysis can quickly and specifically distinguish P.cubensis from other species,which could be utilized for forensic science,medical diagnosis and drug trafficking cases.
基金This study was supported by the National Science Foundation of China(No.30972073,30871766,30471208 and 30070005).
文摘Mushroom poisoning is the main cause of mortality in food poisoning incidents in China.Although some responsible mushroom species have been identified,some were identified inaccuratly.This study investigated and analyzed 102 mushroom poisoning cases in southern China from 1994 to 2012,which involved 852 patients and 183 deaths,with an overall mortality of 21.48%.The results showed that 85.3%of poisoning cases occurred from June to September,and involved 16 species of poisonous mushroom:Amanita species(A.fuliginea,A.exitialis,A.subjunquillea var.alba,A.cf.pseudoporphyria,A.kotohiraensis,A.neoovoidea,A.gymnopus),Galerina sulciceps,Psilocybe samuiensis,Russula subnigricans,R.senecis,R.japonica,Chlorophyllum molybdites,Paxillus involutus,Leucocoprinus cepaestipes and Pulveroboletus ravenelii.Six species(A.subjunquillea var.alba,A.cf.pseudoporphyria,A.gymnopus,R.japonica,Psilocybe samuiensis and Paxillus involutus)are reported for the first time in poisoning reports from China.Psilocybe samuiensis is a newly recorded species in China.The genus Amanita was responsible for 70.49%of fatalities;the main lethal species were A.fuliginea and A.exitialis.Russula subnigricans caused 24.59%of fatalities,and five species showed mortality>20%(A.fuliginea,A.exitialis,A.subjunquillea var.alba,R.subnigricans and Paxillus involutus).Mushroom poisoning symptoms were classified from among the reported clinical symptoms.Seven types of mushroom poisoning symptoms were identified for clinical diagnosis and treatment in China,including gastroenteritis,acute liver failure,acute renal failure,psychoneurological disorder,hemolysis,rhabdomyolysis and photosensitive dermatitis.
