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. 2002 Jul;40(7):2357–2362. doi: 10.1128/JCM.40.7.2357-2362.2002

TABLE 1.

Summary of in-house PCR methods for detection of C. pneumoniae

Variable Laboratory site
A B C D E
PCR format Single Nested Single Nested Nested
Target gene 16S rRNA ompA Cloned PstI ompA ompA
Primers CPNA-CPNB CP1-CP2, CPC-CPD HL1-HR1 CP1-CP2, CPC-CPD CP1-CP2, CPC-CPD
Amplicon size (bp) 463 207 474 207 207
No. of replicates tested     1     3     4     2 1 or 2
Criterion for positivity 1/1 1/3 1/4 1/2 1/1 or 1/2
Size of PCR mixture (μl) 50 50 50 50 100
Vol of original specimen used in PCR (μl) 15 10 0.1, 0.5, 1, 5 10 10
Vol of amplicon transferred in nested PCR (μl) N/Aa     2.5b N/A      5c 10d
Detection method AGEe AGE AGE and hybridization AGE AGE
Reference 7 23 5 23 23
a

N/A, not applicable.

b

The amplicon was diluted 1:10, and 2.5 μl of the dilution was added to 25 μl of the round 2 reaction mixture.

c

The amplicon was diluted 1:2, and 5 μl of the dilution was added to 50 μl of the round 2 reaction mixture.

d

Ten microliters of undiluted amplicon was added to 100 μl of the round 2 reaction mixture.

e

AGE, agarose gel electrophoresis.