Table 4.
Summary of recent multiplex PCR assays for the detection of M. pneumoniae published since 2003 and previously validated assays used as comparators
| Reference | Assay type | Detection format | Target gene (bp) | PCR assay used as a comparator for the new assay | Non-PCR comparator test | Specimens tested for the validation of sensitivity and/or specificity |
|---|---|---|---|---|---|---|
| 2004 [84] | MX-PCR | Microchip electrophoresis | 16S rRNA gene(88) | Mono-assay | Serology |
M. pneumoniae dilution series, various bacterial species, clinical specimens Pathogens targeted: M. pneumoniae, Chlamydophila pneumoniae, and Legionella pneumophila |
| 2005 [85] |
Mass Tag MX-PCR |
Masscode Tag | NS | ND | ND |
DNA dilutions, M. pneumoniae-negative clinical specimens Pathogens targeted: influenza A and B, RSV A and B, metapneumovirus, SARS, coronavirus OC43 and 229E, parainfluenza 1-3, C. pneumoniae, M. pneumoniae, L. pneumophila, enterovirus, adenovirus |
| 2005 [57] |
MX-PCR Chlamylege |
Hybridization | P1 gene (298) | [86] | Serology |
Various bacterial species, bacterial dilutions, clinical specimens with known status, clinical specimens with unknown status Pathogens targeted: C. pneumoniae, M. pneumoniae, Legionella spp. |
| 2005 [56] |
MX-PCR Pneumoplex |
Real-time | 16S rRNA gene | NS | ND |
Various bacterial species, dilutions of recombinant DNA, dilutions of organisms, spiked BALs Pathogens targeted: M. pneumoniae, C. pneumoniae, L. pneumophila, L. micdadei, Bordetella pertussis |
| 2005 [87] | MX-PCR | Agarose gel electrophoresis | P1 gene (360) | [72] | ND |
Various bacterial specimens, dilutions of organisms, clinical specimens with known M. pneumoniae status by PCR Pathogens targeted: M. pneumoniae, C. pneumoniae, L. pneumophila, B. pertussis |
| 2005 [88] | MX-PCR | Agarose gelelectrophoresis | P1 gene (483) | ND | ND |
Various bacterial species, serial dilutions of DNA, clinical respiratory specimens Pathogens targeted: Streptococcus pneumoniae, Haemophilus influenzae, M. pneumoniae, C. pneumoniae |
| 2007 [89] | MX-PCR | Agarose gelelectrophoresis | P1 gene (225) | M.pneumoniae OligoDetect PCR kit, P1-based in-house PCR | ND |
Extracts from samples known to be positive for some common respiratory bacterial pathogens, clinical specimens from children with RTIs Pathogens targeted: M. pneumoniae, C. pneumoniae, B. pertussis, B. parapertussis |
| 2007 [90] | Nested MX-PCR | Agarose gel electrophoresis | P1 gene (343, 160) | ND | ND |
DNA dilutions, clinical specimens Pathogens targeted: M. pneumoniae, C. pneumoniae, L. pneumophila, adenovirus |
| 2007 [55] | MX-PCR | Resequencing microarray | NS | ND | ND |
Various bacterial species, M. pneumoniae cells, DNA dilutions, archived throat swabs Pathogens targeted: adenovirus, Bacillus anthracis, C. pneumoniae, influenza virus A and B, Francisella tularensis, coronavirus 229 E and OC43, rhinovirus, Lassa virus, M. pneumoniae, parainfluenza virus 1, 3, RSV A and B, S. pneumoniae, S. pyogenes, vaccinia virus, Yersinia pestis, Ebola virus, Variola major virus |
| 2007 [91] | MX-PCR | Microarray with electrochemical detection | dnaK gene (654), pdhA gene (284), tuf gene (604) | ND | ND |
Various bacterial and viral species, dilution series The pdhA primers and probes were found to be the most sensitive combination Pathogens targeted: B. pertussis, S. pyogenes, C. pneumoniae, M. pneumoniae, adenovirus, coronavirus OC43, 229E, and HK, influenza A and B, parainfluenza types 1, 2, and 3, RSV |
| 2008 [58] |
MX-PCR ResPlex |
Luminex technology | ATPase (NS) | NS | ND |
M. pneumoniae type I and II , >110 strains of other bacterial species found in the respiratory tract or related spp., 10-fold serial DNA dilutions, 49 NPS with known M. pneumoniae status by PCR Pathogens targeted: S. pneumoniae, Neisseria meningitidis, H. influenzae, L. pneumophila, M. pneumoniae, C. pneumoniae |
| 2008 [92] | MX-PCR | Molecular beacons | P1 gene (158) | [93] | ND |
Reference strains of common bacterial respiratory pathogens or related species, serial dilutions of DNA, spiked samples, samples with known status by PCR Pathogens targeted: M. pneumoniae and C. pneumoniae |
| 2008 [94] | MX NASBA | Molecular beacons | 16S rRNA | [81, 73] | Culture, serology |
Various bacterial species, bacterial dilutions, dilutions of wild-type M. pneumoniae 16S rRNA generated in vitro, spiked specimens, specimens with known status by PCR Pathogens targeted: M. pneumoniae, C. pneumoniae, Legionella spp. |
| 2008 [95] | MX-PCR | Reverse line blot hybridization | 16S-23S rRNA spacer (93) | [60] | ND |
12 reference strains and 63 clinical isolates of common bacterial respiratory pathogens, 10-fold serial DNA dilutions, 100 NPAs from children with CAP Pathogens targeted: Staphylococcus aureus, S. pneumoniae, S. pyogenes, Moraxella catarrhalis, H. influenzae, B. pertussis, Klebsiella pneumoniae, L. pneumophila, Mycobacterium tuberculosis, C. pneumoniae, M. pneumoniae |
| 2008 [96] | MX-PCR | Enzyme hybridization or electronic microarray detection | P1 gene (299) | ND | ND |
Various bacterial and viral species, bacterial dilutions, spiked clinical specimens, clinical specimens from carriage study, re-analysis of samples with discrepant results Pathogens targeted: influenza virus A, influenza virus B, RSV A and B, M. pneumoniae, C. pneumoniae, L. pneumophila, L. micdadei, B. pertussis, S. aureus, S. pneumoniae |
A-test: passive agglutination-test; MX-PCR: multiplex PCR; NASBA: nucleic acid sequence-based amplification; NPA: nasopharyngeal aspirate; NS: not specified; Pneumoplex (GenProbe Prodesse Inc., Waukesha, WI, USA); ResPlex (Qiagen GmbH, Valencia,CA, USA); M. pneumoniae OligoDetect PCR kit (Millipore/Chemicon, Eugene, OR, USA)