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Table 1 Amplification conditions, oligonucleotide combinations, sequence and amplification fragment of multiplex-PCR for K. variicola identification

From: Development of a Multiplex-PCR probe system for the proper identification of Klebsiella variicola

Amplification conditions a

Name of combination primers

Shared unique genes, oligonucleotides and sequence (5′- 3′) of each bacterial specie

K. pneumoniae

Amplification fragment (bp)

K. variicola

Amplification fragment (bp)

Klebsiella spp.

Amplification fragment (bp)

  

phosphohydrolase

 

phosphoglycerate mutase

 

phosphopentane phosphatase (mtnC)

 

1

M-PCR-1

KP888-F: AAGCAAGCCAGAACAGAAAG

888

KV770-F: TCCCGAGGTTCACATTTCC

449

KmtnC-F: CCGCCGACCTTATCACTAC

340

  

KP888-R: ACTTCGGTTTTATCCAGGTC

 

KV770-R: AGCGGGTGAACGTCGATAC

 

KmtnC-R: AGCGGGTGAACGTCGATAC

 
  

transferase (yphG)

 

N-acetyltransferase

 

phosphopentane phosphatase (mtnC)

 

1

M-PCR-2

KP878-F: ACCGATAACCAGCCTGACTT

878

KV1615-F: ACACAACATTTCAGGCGGCT

499

KmtnC-F: CCGCCGACCTTATCACTAC

340

  

KP878-R: CTTTCTTCTGCCCACTGTTG

 

KV1615-R: GGGCGTGGCTTTTTTCATCG

 

KmtnC-R: AGCGGGTGAACGTCGATAC

 
  

phosphohydrolase

 

thiopurine S-methyltransferase

 

phosphopentane phosphatase (mtnC)

 

2

M-PCR-3

KP888-F: AAGCAAGCCAGAACAGAAAG

888

KV1000-F: CTGGGATGTGGCAATGGTG

438

KmtnC-F: CCGCCGACCTTATCACTAC

340

  

KP888-R: ACTTCGGTTTTATCCAGGTC

 

KV1000-F: AAACTGCGCCTGCTGTATC

 

KmtnC-R: AGCGGGTGAACGTCGATAC

 
  1. aMultiplex-PCR conditions used under the oligonucleotides combinations. 1: 5pmol/reaction of K. variicola and Klebsiella spp, 25pmol/reaction of K. pneumoniae; 2: 25 pmol/reaction of K. pneumoniae, 5 pmol/reaction of K. variicola and 1 pmol/reaction of Klebsiella spp.