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US 2010/0167272 A1Published application
Method for Detecting and Analyzing Pathogens in a Sample
AssigneeUniversita' Degli Studi Di Firenze
InventorsChiara Azzari, Massimo Resti, Maria Moriondo
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Abstract
The present invention relates to a method and kits thereof for detecting the presence and/or the specific serogroup of a prokaryote selected from the group consisting of & pneumionae, N. meningitidis, H. influenzae, Adenovirus, Klebsiella Pneumonite, Lysteria monocytogenes, Escherichia coli and Streptococcus agalactiae in a sample taken from a human being, through amplification of specific target regions.
Claims22
Method for detecting the presence and/or the serogroup of a pathogen selected from the group consisting of N. meningitidis, H. influenzae, S. pneumoniae or Adenovirus in a biological sample, comprising the following steps:
- a) incubating a first aliquot of the sample under conditions such as to enable the amplification and the revelation of specific target regions of the genoma of said pathogens, if present in the sample, wherein the target regions are comprised in:
- SEQ ID 119 of the ctrA gene of N. meningitidis, and
- SEQ ID 120 of the P2 gene or SEQ ID 121 of the bex gene of H. influenzae and
- SEQ ID 122 of the lyt gene or SEQ ID 123 of the ply gene of S. pneumoniae and
- SEQ ID 124 of Adenovirus;
- and/or,
- b) if the sample is postive for N. meningitidis, incubating a second aliquot of the sample under conditions such as to enable the aplification of specific serotyping target regions of the genoma of N. meningitidis, wherein the target regions are comprised in:
- SEQ ID 125 for serotype B of N. meningitidis,
- SEQ ID 126 for serotype C of N. meningitidis,
- SEQ ID 127 for serotype W135 of N. meningitidis,
- SEQ ID 128 for serotype W of N. meningitidis,
- SEQ ID 129 for serotype Y of N. meningitidis,
- SEQ ID 130 for serotype A of N. meningitidis,
- c) revealing the amplification;
- b′) if the sample is positive for H. influenzae, incubating a second aliquot of the sample under conditions such as to enable the amplification of region SEQ ID 131 of the genoma of H. influenzae for revealing capsulated H. influenzae, and
- c′) revealing the amplification;
- d′) if the sample is positive for the revelation of capsulated H. influenzae, incubating a third aliquot of the sample under conditions such as to enable the amplification of specific serotyping target regions of the genoma of H. influenzae wherein the target regions are comprised in:
- SEQ ID 132 for the revelation of H. influenzae that are productors of beta-lactamase
- SEQ ID 133 for the revelation of H. influenzae serotypes a, b, c, d, e, f;
- SEQ ID 134 for the revelation of H. influenzae B type capsulated serotype;
- b″) if the sample is positive for S. pneumonite, incubating a second aliquot of the sample under conditions such as to enable the amplification of specific serotyping target regions of the genoma of S. pneumoniae, wherein the target regions are comprised in:
- SEQ ID 135 for serotype 19F of S. pneumoniae
- SEQ ID 136 for serotype 22F of S. pneumoniae
- SEQ ID 137 for serotype 3 of S. pneumoniae
- SEQ ID 138 for serotype 6 of S. pneumoniae
- SEQ ID 139 for serotype 19A of S. pneumoniae
- SEQ ID 140 for serotype 9v of S. pneumoniae
- SEQ ID 141 for serotype 4 of S. pneumoniae
- SEQ ID 142 for serotype 14 of S. pneumoniae
- SEQ ID 143 for serotype 12f of S. pneumoniae
- SEQ ID 144 for serotype 7f of S. pneumoniae
- SEQ ID 145 for serotype 11a of S. pneumoniae
- SEQ ID 146 for serotype 33f of S. pneumoniae
- SEQ ID 147 for serotype 16f of S. pneumoniae
- SEQ ID 148 for serotype 35b of S. pneumoniae
- SEQ ID 149 for serotype 18f of S. pneumoniae
- SEQ ID 150 for serotype 38 of S. pneumoniae
- SEQ ID 151 for serotype 31 of S. pneumoniae
- SEQ ID 152 for serotype 15c of S. pneumoniae
- SEQ ID 153 for serotype 8 of S. pneumoniae
- SEQ ID 154 for serotype 10A of S. pneumoniae
- SEQ ID 155 for serotype 35f of S. pneumoniae
- SEQ ID 156 for serotype 34 of S. pneumonite
- SEQ ID 157 for serotype 1 of S. pneumoniae
- SEQ ID 158 for serotype 17f of S. pneumoniae
- SEQ ID 159 for serotype 20 of S. pneumoniae
- SEQ ID 160 for serotype 15a of S. pneumoniae
- SEQ ID 161 for serotype 7c of S. pneumoniae
- SEQ ID 162 for serotype 18f of S. pneumoniae
- SEQ ID 163 for serotype 5 of S. pneumoniae
- SEQ ID 164 for serotype 23F of S. pneumoniae
- c″) highlighting the amplification;
- d″) if the sample is positive for serotype 6 of S. pneumoniae, incubating a third aliquot of the sample under conditions such as to enable the amplification of the regions SEQ ID 165 for serotype 6a or SEQ ID 166 for serotype 6b of S. pneumoniae
- e″) highlighting the amplification.
51 more elements
Method for detecting the presence and/or the serogroup of a pathogen selected from the group consisting of Klebsiella pneumoniae, Lysteria monocytogenes, E. coli, S. agalactiae in a biological sample comprising the following steps:
- a) incubating an aliquot of the sample under conditions such as to enable the amplification and revelation of specific target regions of the genoma of said pathogens, if present in the sample, wherein the target regions are comprised in:
- SEQ ID 167 of the phoE gene of Klebsiella pneumoniae,
- SEQ ID 168 of the iap gene of Lysteria monocytogenes,
- SEQ ID 169 of the uidA gene of E. coli,
- SEQ ID 170 of the sip gene of S. agalactiae.
2 more elements
Dependencies are inferred from the claim text.