KS I ISO 10705-1-2008 Water quality－Detection and enumeration of bacteriophages－Part 1：Enumeration of F-specific RNA bacteriophages《水质 噬菌体的检查和计数 第1部分 F 此RNA噬菌体的计数》.pdf

1、 KSKSKSKSKSKSKSK KSKSKS KSKSK KSKS KSK KS KS I ISO 10705 1 1： F- RNA KS I ISO 10705 1:2008 2008 11 10 http:/www.kats.go.krKS I ISO 10705 1:2008 : ( ) ( ) ( ) : (http:/www.standard.go.kr) ： ：1998 12 24 ：2008 11 10 2008-0751 ： ： ( 02-509-7266) (http:/www.kats.go.kr). 10 5 , . KS I ISO 10705 1:2008 1：

2、F- RNA Water quality Detection and enumeration of bacteriophagesPart 1： Enumeration of F-specific RNA bacteriophages 1995 1 ISO 10705 1, Water quality Detection and enumeration of bacteriophages Part 1： Enumeration of F-specific RNA bacteriophages , . 1 F (RNA) . , , . KS M ISO 10705 . . F RNA , . F

3、 RNA . , , A , . 2 . . ( ) . KS M ISO 3696, KS M ISO 5667 1, ( ) KS M ISO 5667 2, 2： KS M ISO 5667 4, 4： KS M ISO 8199, ISO 6887： 1983, Microbiology General guidance for the preparation of dilutions for microbiological KS I ISO 10705 1:2008 2 examination 3 . 3.1 F RNA F (sex-pili) (plaque) , 40( 400

4、) g/mL RN 4 . . . RN . (Cpfp) . 5 , . F RNA . . . 6 , 6.1 , A . KS M ISO 8199 . . , KS M ISO 3696 . 6.2 A.8 . KS I ISO 10705 1:2008 3 6.3 6.3.1 RNase 50 units/mg(Kunitz) . 6.3.2 (130 g： 9 mm) (100 g： 9 mm) . 6.3.3 870 g/L . 6.4 WG49, 3 Nalr(F42 lac:Tn5), NCTC 12484 MS2, NCTC 12487 ATCC 15597 B1 K 12

5、 Hfr, NCTC 12486 ATCC 23631 1 NCTC National Collection of Type Cultures, 61 Colindale Avenue, London NW9 6HT, England . ATCC American Type Culture Collection, 12301 Parklawn Drive, Rockville, Maryland, USA . 7 . 7.1 , KS M ISO 8199 . 7.2 (371) . 7.3 (371) , (10010) min 1 . 7.4 (451) . 7.5 KS I ISO 1

6、0705 1:2008 4 7.6 pH 7.7 ( ) . 7.8 ( 205) . 7.9 ( 7010) . 7.10 1 cm (7.17 ) . 10 nm 500 nm650 nm . KS M ISO 8199 , 1 . 7.11 9 cm 15 cm . 7.12 1 mL, 5 mL 10 mL . 7.13 . 7.14 . 7.15 . 7.16 250 mL300 mL . 7.17 KS I ISO 10705 1:2008 5 1 cm (7.10 ) , 250 mL300 mL ( 1 ) 7.18 0.2 m 7.19 1.5 mL2 mL . 1 8 KS

7、 M ISO 5667 1, KS M ISO 5667 2, KS M ISO 5667 3 KS M ISO 8199 . 9 9.1 WG49 K12 Hfr , 2 . . KS I ISO 10705 1:2008 6 2 WG49 , 9.1.1 TYGB(A.1) (rehydrate) . 300 mL (7.16) TYGB 50 mL . (10010) min 1 (371) (182) h . (A.6) 10 mL . (7.19) 1.2 mL ( 7010) . 2 . 9.1.2 (9.1.1) , (A.7) . (371) (182) h . TYGB(A.

8、1 ) 50 mL 300 mL (7.16) . 3 5 , TYGB . (10010) min 1 (371) (51) h . , 20 %(V/V) 70 2 (9.1.2) 2 (10.1) (9.1.1) (log-phase) 20 %(V/V) (stock) 70 (9.1.1) (9.3)KS I ISO 10705 1:2008 7 (A.6) 10 mL . (7.19) 1.2 mL ( 7010) 2 . 9.3 . 3 . 4 . , . . 9.2 WG49 . TYGB(A.1) 50 mL (7.17) 0 . (7.16) 0 . 0.5 mL . (1

9、0010) min 1 3 (371) . 30 1 mL . 10 6 , 10 4, 10 5 10 6 0.1 mL TYGA (A.2) (371) (242) h . 30300 , mL cfp ( KS M ISO 8199 ). 5 . , . 9.3 WG49 9.2 . t 0 t 3 (A.7) (371) (242) h . 30300 , . t 0 t 3 10 2 0.1 mL (Nal) , (Km) (371) (242) h . . . 0 TYGA(9.2) ： 0.5cfp/mL3107 cfp/mL 3 TYGA(9.2) ： 7cfp/mL40107

10、 cfp/mL ( ) 8 % Nal ： . Km ： 20 mm KS I ISO 10705 1:2008 8 6 . Km . F RNA . C MS2 (42) . 10 , 10.1 . K 12 Hfr . (42) . , 100 pfp/mL MS2 (A.8) 100 mL1 000 mL . (5 g/L) . (7.19) 1.2 mL , ( 205) ( 705) . 4 10.1 K 12 Hfr WG49 1 mL . , WG49 . 80 % WG49 . 10 10.1 . TYGB(A.1) 50 mL (7.17) . 9.2 0 . 0.5 mL

11、. (10010) min 1 (371) . 30 . 108cfu/mL(9.2 ) . 2 . 7 F , F . ssTYGA(A.3) , 44 50 (A.1) (0.5 mL/50 mL) , 2.5 mL (451) . ( ) 1 mL . . 1 mL , TYGA (A.2) 9 cm . , , (371) (182) h . 8 6 ( 4 ) . 9 . . KS I ISO 10705 1:2008 9 10 . 4 . 10.2 100 g /mL ssTYGA(A.3) . 10 . ssTYGA (A.4) (0.2 mL/50 mL) TYGA . 10.

12、3 10.1 40 g/mL ssTYGA RN (A.5) ( , RN 100 L ssTYGA 2.5 mL ). 11 . a) b) NRNase/N(11. ) 10 % c) NRNase/N 10 % d) ( ) 12 RNA RNase 40 g/mL RN 400 g/mL . 10.4 (phage) 10.1 . ssTYGA 10 mL, 1 mL 5 mL 14 cm TYGA 50 mL . 13 10 20 , 50 mL 100 mL 1 pfu . . 10.5 MS2(9.3 ) . . 100 pfp/mL (5 g/L) , ( 205) ( 705

13、) . RNA . 14 . KS I ISO 10705 1:2008 10 ( .) 9.1.2 . 11 30 300 . , 1 mL F RNA ( ) . FnNNC =RNasepfp Cpfp ： mL F RNA N ： 10.1, 10.2 10.4 WG49 NRNase ： 10.3 RN WG49 n ： F ： ( ) (10.4 ) 12 . a) b) c) N NRNase d) 11. e) 1 5 KS I ISO 10705 1:2008 11 A ( ) , A.1 (TVGB) 10 g1 g8 g1 000 mL . 25 (7.20.1) pH

14、. 200 mL , (1211) 15 . (42) 6 . (2 ) 3 g10 g100 mL . 0.22 m . (42) 6 . 200 mL2 mL . (42) 6 . A.2 (TYGA) 10 g1 g8 g1220 g1)1 000 mL1) . KS I ISO 10705 1:2008 12 . 25 (7.20.1) pH . 200 mL (1211) 15 . (42) 6 . (A.1) 200 mL2 mL 45 50 . . 9 cm 20 mL 14 cm 50 mL (42) 6 . A.3 (ssTYGA) A.2 , (6 g10 g) . ssT

15、YGA . 50 mL . A.4 (1 mol/L) 250 mg2 mL8 mL . 0.22 m . (42) 8 ( 202) 6 . A.5 RN ( ) RN 100 mg100 mL100 10 RNase . 0.5 mL 20 1 . . A.6 ( ) (870 g/L) 100 mL 20 mL (1211) 15 . 1 . KS I ISO 10705 1:2008 13 A.7 (McConkey) 20.0 g 10.0 g 5.0 g 75 mg 1220 g 1 000 mL . 25 (7.40.1) pH . 200 mL (1211) 15 . 45 5

16、0 , 9 cm 20 mL . (42) 6 . A.8 1.0 g8.5 g1 000 mL 950 mL . (7.00.1) (1 mol/L) pH . 1 000 mL , . (1211) 15 . 6 . KS I ISO 10705 1:2008 14 B ( ) F RNA F RNA 21 nm30 nm , RNA ( ) . E Leviviridae (family) . 2 (genera) . MS2 Levivirus Q2 Allolevivirus . K12 F , F (sex) . F . RN , F RNA Inoviridae (family) F DNA . KS I ISO 10705 1:2008 15 C ( ) MS2 . . TYGB 25 mL 300 mL , ( K 12 Hfr, NCTC 12486) . (10010) min 1 (371) (182) h . 300 mL TYGB 25 mL 35 37 , 0.25 mL . 90 . MS2 107 pfp/mL . 4 h5 h . (CHCl3) 2.5 mL , (42) . , 3 000 g 20