Estimating included animal species in mixed crude drugs derived from animals using massively parallel sequencing

Abstract We developed a method that can detect each animal species of origin for crude drugs derived from multiple animal species based on massively parallel sequencing analysis of mitochondrial genes. The crude drugs derived from animals investigated in this study were Cervi Parvum Cornu and Trogop...

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Autores principales:	Hiroaki Nakanishi, Katsumi Yoneyama, Masaaki Hara, Aya Takada, Kazuyuki Saito
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/24913b7cf6654cf695c81ed8fb99d344
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spelling	oai:doaj.org-article:24913b7cf6654cf695c81ed8fb99d3442021-12-02T13:17:42ZEstimating included animal species in mixed crude drugs derived from animals using massively parallel sequencing10.1038/s41598-021-85803-42045-2322https://doaj.org/article/24913b7cf6654cf695c81ed8fb99d3442021-03-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-85803-4https://doaj.org/toc/2045-2322Abstract We developed a method that can detect each animal species of origin for crude drugs derived from multiple animal species based on massively parallel sequencing analysis of mitochondrial genes. The crude drugs derived from animals investigated in this study were Cervi Parvum Cornu and Trogopterorum feces, which are derived from a mix of different animal species, two chopped cicada sloughs, and two commercial Kampo drugs. The mitochondrial 12S rRNA, 16S rRNA, and cytochrome oxidase subunit I gene regions were amplified and sequenced using MiSeq. The ratios of haplotype to total number of sequences reads were calculated after sequence extraction and trimming. Haplotypes that exceeded the threshold were defined as positive haplotypes, which were compared with all available sequences using BLAST. In the Cervi Parvum Cornu and Trogopterorum feces samples, the haplotype ratios corresponded roughly to the mixture ratios, although there was a slight difference from mixture ratios depending on the gene examined. This method could also roughly estimate the compositions of chopped cicada sloughs and Kampo drugs. This analysis, whereby the sequences of several genes are elucidated, is better for identifying the included animal species. This method should be useful for quality control of crude drugs and Kampo drugs.Hiroaki NakanishiKatsumi YoneyamaMasaaki HaraAya TakadaKazuyuki SaitoNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-8 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Hiroaki Nakanishi Katsumi Yoneyama Masaaki Hara Aya Takada Kazuyuki Saito Estimating included animal species in mixed crude drugs derived from animals using massively parallel sequencing
description	Abstract We developed a method that can detect each animal species of origin for crude drugs derived from multiple animal species based on massively parallel sequencing analysis of mitochondrial genes. The crude drugs derived from animals investigated in this study were Cervi Parvum Cornu and Trogopterorum feces, which are derived from a mix of different animal species, two chopped cicada sloughs, and two commercial Kampo drugs. The mitochondrial 12S rRNA, 16S rRNA, and cytochrome oxidase subunit I gene regions were amplified and sequenced using MiSeq. The ratios of haplotype to total number of sequences reads were calculated after sequence extraction and trimming. Haplotypes that exceeded the threshold were defined as positive haplotypes, which were compared with all available sequences using BLAST. In the Cervi Parvum Cornu and Trogopterorum feces samples, the haplotype ratios corresponded roughly to the mixture ratios, although there was a slight difference from mixture ratios depending on the gene examined. This method could also roughly estimate the compositions of chopped cicada sloughs and Kampo drugs. This analysis, whereby the sequences of several genes are elucidated, is better for identifying the included animal species. This method should be useful for quality control of crude drugs and Kampo drugs.
format	article
author	Hiroaki Nakanishi Katsumi Yoneyama Masaaki Hara Aya Takada Kazuyuki Saito
author_facet	Hiroaki Nakanishi Katsumi Yoneyama Masaaki Hara Aya Takada Kazuyuki Saito
author_sort	Hiroaki Nakanishi
title	Estimating included animal species in mixed crude drugs derived from animals using massively parallel sequencing
title_short	Estimating included animal species in mixed crude drugs derived from animals using massively parallel sequencing
title_full	Estimating included animal species in mixed crude drugs derived from animals using massively parallel sequencing
title_fullStr	Estimating included animal species in mixed crude drugs derived from animals using massively parallel sequencing
title_full_unstemmed	Estimating included animal species in mixed crude drugs derived from animals using massively parallel sequencing
title_sort	estimating included animal species in mixed crude drugs derived from animals using massively parallel sequencing
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/24913b7cf6654cf695c81ed8fb99d344
work_keys_str_mv	AT hiroakinakanishi estimatingincludedanimalspeciesinmixedcrudedrugsderivedfromanimalsusingmassivelyparallelsequencing AT katsumiyoneyama estimatingincludedanimalspeciesinmixedcrudedrugsderivedfromanimalsusingmassivelyparallelsequencing AT masaakihara estimatingincludedanimalspeciesinmixedcrudedrugsderivedfromanimalsusingmassivelyparallelsequencing AT ayatakada estimatingincludedanimalspeciesinmixedcrudedrugsderivedfromanimalsusingmassivelyparallelsequencing AT kazuyukisaito estimatingincludedanimalspeciesinmixedcrudedrugsderivedfromanimalsusingmassivelyparallelsequencing
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Estimating included animal species in mixed crude drugs derived from animals using massively parallel sequencing

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