Hanyoyin PCR da gurɓataccen iska a cikin dakunan gwaje-gwaje na acid nucleic kamar ɓangarori biyu ne na tsabar kuɗi.Za mu iya zaɓar samun shi ne kawai ko a'a, amma ba za mu iya zaɓar ko muna so ko mu kashe shi ba.

1. Nunawa na cire DNA

Don cimma nasarar kawar da gurɓataccen iska na nucleic acid aerosol, da farko ya zama dole a tantance masu cire DNA waɗanda zasu iya cire acid nucleic a cikin yanayin ruwa.Domin babu masu cire DNA da yawa waɗanda suke aiki da gaske.Don hanyar gwaji, da fatan za a koma zuwa: Mai cire DNA ba zai iya zama "kusurwar sirri" na dakin gwaje-gwaje ba!

A cikin wannan gwaji, 100 kofe / μL (CT kusan 31) na ASFV plasmid da DNA cirewa da aka ƙididdige su ta PCR na dijital an gauraye su a daidai adadin, sa'an nan kuma aka mayar da martani a cikin zafin jiki na 10min, 20min da 30min bi da bi.Bayan hakar acid nucleic, an aiwatar da haɓaka qPCR.An kwatanta ingantaccen iko mai gauraye da plasmid da ruwa.Tun da ba a kammala gwajin a lokaci guda ba, za a iya samun wata karkata tsakanin sakamakon, amma hakan bai shafi ƙarshen gwajin ba.Ya zuwa yanzu, na kimanta samfuran cire DNA na kasuwanci guda 10.Na'urar No. 1, No. 6 da No. 8 ne kawai samfuran zasu iya lalata DNA plasmid yadda ya kamata a cikin yanayin ruwa.Sauran samfurori ba su da wani tasiri.

Tebur 1 Tasirin kawar da acid nucleic na mai cire DNA na kasuwanci

2. Gwajin tasirin kawar da DNA na magungunan kashe kwayoyin cuta da aka saba amfani da su

1. Binciken magungunan kashe kwayoyin cuta da aka saba amfani da su

Akwai nau'ikan magungunan kashe kwayoyin cuta da yawa da aka saba amfani da su a dakunan gwaje-gwaje: aldehydes, phenols, alcohols, salts ammonium quaternary, peroxides, shirye-shiryen chlorine, da acid da tushe.An tabbatar da illar cututtukan da waɗannan magungunan kashe kwayoyin cuta ke yi ta hanyar gwaje-gwaje, amma babu isassun bayanan gwaji kan lalata tasirin acid nucleic.Dakunan gwaje-gwaje na gwajin Nucleic acid suna buƙatar maganin kashe kwayoyin cuta wanda ba wai kawai yana da kyakkyawan tasirin kashe ƙwayoyin cuta ba, har ma yana iya lalata DNA.Sakamakon gwajin ya nuna cewa shirye-shiryen chlorine guda biyu ne kawai na maganin kashe kwayoyin cuta 84 da trichloroisocyanuric acid da 1 M hydrochloric acid sun cika buƙatun.Tun da 1M hydrochloric acid yana da lalata sosai, ana ba da shawarar cewa a yi amfani da shirye-shiryen chlorine a matsayin zubar da shara a dakunan gwaje-gwaje na gwajin nucleic acid.Koyaya, shirye-shiryen chlorine suna lalata ga karafa kuma ba za a iya amfani da su don lalata kayan aiki da kayan aiki ba.

Tebur na 2 Sakamakon cirewar DNA na nau'ikan magungunan kashe qwari

2. Mafi ƙarancin tasiri na shirye-shiryen chlorine

Abubuwan da ake amfani da su na chlorine suna da tasiri mai ƙarfi na lalata DNA, amma saboda lalatawar ƙarfe da haushi, ana iya amfani da su don tsaftace ƙasa, wuraren da ba na ƙarfe ba, tukwici mai jiƙa, bututun centrifuge da sauran abubuwan gwaji.

Bisa ga "Sabuwar Cutar Kwayoyin Cutar Kwayoyin Cutar Kwayoyin Cutar Kwayoyin cuta": Ƙaƙƙarfan ƙazantawa (jinin marasa lafiya, ɓoyewa da amai) akwai sinadarin chlorine 5g/L-10g/L mai ɗauke da chlorine;benaye, bango, da saman abu suna amfani da sinadarin chlorine 1g/L Chlorine: Tufafi, kwanciya da sauran kayan yadi ana fara jiƙa a cikin maganin chlorine tare da 0.5g/L akwai chlorine na mintuna 30, sannan a tsaftace kamar yadda aka saba.

Tebura 3 Tasirin kawar da DNA na ma'auni daban-daban na maganin chlorine

Sakamakon gwajin ya nuna cewa: lokacin da tasirin chlorine mai inganci ya fi ko daidai da 1.2 g/L, maganin da ke ɗauke da chlorine zai iya lalata plasmid 100 kwafi/μL gaba ɗaya idan aka yi amfani da shi na mintuna 5.Lokacin yin aiki na mintuna 10, maganin kashe ƙwayoyin cuta guda 84 tare da ingantaccen ƙwayar chlorine mafi girma ko daidai da 0.6 g/L na iya lalata kwafin 100/μL na plasmids gaba ɗaya.

3. Gwajin kawar da iska da aka gurbata da aerosol na nucleic acid

Yadda za a tsaftace iskar da aka gurbata ta hanyar aerosols na nucleic acid?Yawancin tunani shine ƙirƙirar yanayi mai gurɓataccen iska sannan a tsaftace shi, ko kuma tsabtace acid nucleic bayan gurɓata.A gare ni, gwajin da ba za a iya maimaita shi ba kuma ba za a iya tantance sakamakon gwajin da ƙididdigewa ba ba shi da ma'ana, don haka na ari wasu hanyoyin kawar da iska.Duba "GB27948-2020 Gabaɗaya Bukatun don Magungunan Iskar iska" da "Ka'idodin Fasaha na Disinfection"

1. Gwaji kayan

1.1 Mai cire DNA: A cikin wannan gwaji, an zaɓi masu cire DNA guda biyu na kasuwanci 6 da 8 a cikin Table 1.

1.2 Gwajin plasmid: Yin la'akari da cewa ƙimar CT na mafi yawan gurɓataccen dakin gwaje-gwaje zai kasance mafi girma fiye da 30, ƙaddamarwar ASFV gene plasmid da aka yi amfani da shi a wannan lokacin shine kimanin kwafi 100 / μL, kuma PCR CT mai kyalli yana da kusan 31.07.Ƙara plasmid ɗin zuwa swab ɗin auduga kuma ya bushe.

1.3 Gwaji kayan aiki: Bilingkehan KVBOX disinfection kayan, wanda aka kirki goyon bayan Shenzhen Runlian Environmental Technology Co., Ltd., yana da kyau kwarai aikin disinfection iska tare da hydrogen peroxide disinfectant, da kuma iya yadda ya kamata kashe spores da sauran microorganisms.

1.4 Wurin gwaji: Rufaffen taga canja wuri na kusan mita 0.1 cubic.

2. Hanyar gwaji

Adadin maganin da aka saita akan injin disinfection shine 10ml, kuma maida hankali shine 100ml/m3, wanda ya fi girma fiye da "GB27948-2020 Gabaɗaya Bukatun Don Magungunan Iska": lokacin amfani da maganin feshin aerosol, adadin maganin ya kamata ya zama ≤10 ml/m3