Evaluation on multiplex PCR for detecting the common serotypes of one kind of Streptococcus pneumoniae

doi:10.3969/j.issn.1673-8640.2015.06.014

Abstract

Abstract: Objective

To evaluate the performance of multiplex polymerase chain reaction (PCR) for detecting the common serotypes of one kind of Streptococcus pneumoniae.

Methods

A total of 50 isolates of Streptococcus pneumoniae were analyzed by multiplex PCR with traditional quellung reaction as reference method for detecting the common serotypes of Streptococcus pneumoniae.

Results

The 50 isolates were typed by traditional quellung reaction with 9 isolates being 19F, 19A, 23F, 6A/B and 14, respectively, and 5 isolates being 15B/C. However, according to the results of multiplex PCR, 8 of 50 isolates were serotype 19F, and there also were 19A (10 isolates), 23F (9 isolates), 6A/B (9 isolates), 14 (10 isolates) and 15B/C (4 isolates), which showed coincidence rate 96%. The value of Kappa was 0.952.

Conclusions

Multiplex PCR has good consistency with traditional method, and it has the advantages of easy, rapid performance and low costing, which could provide the reference for monitoring the serotypes of Streptococcus pneumoniae in clinic.

Key words: Multiplex polymerase chain reaction, Serotype, Streptococcus pneumoniae

CLC Number:

Q503

PAN Fen, ZHAO Shengyuan, QIN Huihong, ZHU Junying, XIAO Shuzhen, ZHANG Hong. Evaluation on multiplex PCR for detecting the common serotypes of one kind of Streptococcus pneumoniae[J]. Laboratory Medicine, 2015, 30(6): 599-602.

Figures/Tables 3

组	P	Q	R	S	T	非疫苗覆盖的血清群/型
A	1^*	1 $8 18 F 、 18 A 、 18 B 、 18 C) *$	4^*	5^*	2^*
B	1 $9 19 F 、 19 A 、 19 B 、 19 C) *$	$6 6 A 、 6 B 、 6 C) *$	3^*	8^*
C	$7 7 F 、 7 A 、 7 B 、 7 C) *$				20^*	24_{(24F、24A、24B)},31,40
D			$9 9 A 、 9 L 、 9 N 、 9 V) *$		1 $1 11 F 、 11 A 、 11 B 、 11 C 、 11 D) *$	16_(16F、16A),36,37
E			1 $2 12 F 、 12 A 、 12 B) *$	1 $0 10 F 、 10 A 、 10 B 、 10 C) *$	3 $3 33 F 、 33 A 、 33 B 、 33 C 、 33 D) *$	21,39
F				1 $7 17 F 、 17 A) *$	2 $2 22 F 、 22 A) *$	27,32_(32F、32A),41_(41F、41A)
G						29,34,35_{(35F、35A、35B、35C)},42,47_(47F、47A)
H	14^*	2 $3 23 F 、 23 A 、 23 B) *$		1 $5 15 F 、 15 A 、 15 B 、 15 C) *$		13,28_(28F、28A)
I						25_(25F、25A),38,43,44,45,46,48

组	P	Q	R	S	T	非疫苗覆盖的血清群/型
A	1^*	1 $8 18 F 、 18 A 、 18 B 、 18 C) *$	4^*	5^*	2^*
B	1 $9 19 F 、 19 A 、 19 B 、 19 C) *$	$6 6 A 、 6 B 、 6 C) *$	3^*	8^*
C	$7 7 F 、 7 A 、 7 B 、 7 C) *$				20^*	24_{(24F、24A、24B)},31,40
D			$9 9 A 、 9 L 、 9 N 、 9 V) *$		1 $1 11 F 、 11 A 、 11 B 、 11 C 、 11 D) *$	16_(16F、16A),36,37
E			1 $2 12 F 、 12 A 、 12 B) *$	1 $0 10 F 、 10 A 、 10 B 、 10 C) *$	3 $3 33 F 、 33 A 、 33 B 、 33 C 、 33 D) *$	21,39
F				1 $7 17 F 、 17 A) *$	2 $2 22 F 、 22 A) *$	27,32_(32F、32A),41_(41F、41A)
G						29,34,35_{(35F、35A、35B、35C)},42,47_(47F、47A)
H	14^*	2 $3 23 F 、 23 A 、 23 B) *$		1 $5 15 F 、 15 A 、 15 B 、 15 C) *$		13,28_(28F、28A)
I						25_(25F、25A),38,43,44,45,46,48

血清型	GenBank编号	引物序列	产物大小(bp)
19F	CR931678	F: GTTAAGATTGCTGATCGATTAATTGATATCC	304
		R: GTAATATGTCTTTAGGGCGTTTATGGCGATAG
19A	CR931675	F: GAGAGATTCATAATCTTGCACTTAGCCA	566
		R: CATAATAGCTACAAATGACTCATCGCC
23F	CR931685	F: GTAACAGTTGCTGTAGAGGGAATTGGCTTTTC	384
		R: CACAACACCTAACACTCGATGGCTATATGATTC
6A/B	CR931639	F: AATTTGTATTTTATTCATGCCTATATCTGG	250
		R: TTAGCGGAGATAATTTAAAATGATGACTA
14	CR931653	F: CTTGGCGCAGGTGTCAGAATTCCCTCTAC
		R: GCCAAAATACTGACAAAGCTAGAATATAGCC
15B/C	CR931664	F: TTGGAATTTTTTAATTAGTGGCTTACCTA
		R: CATCCGCTTATTAATTGAAGTAATCTGAACC
cpsA	AF057294	F: GCAGTACAGCAGTTTGTTGGACTGACC
		R: GAATATTTTCATTATCAGTCCCAGTC

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