Detection of neisseria meningitidis by loop mediated isothermal amplification

Abstract

The invention provides a LAMP assay for detection of meningococcal disease, the test comprising at least one nucleic acid primer set capable of detecting Neisseria meningitides in a LAMP based molecular test, the primer set being chosen from the primer sets listed in Table 1 as LAMP SETS 1 to 12 comprising SEQUENCE IDs from ID: 1 to ID: 69. Each assay consists of a primer set including of one pair of forward (HP) and reverse (BIP) inner primers, forward (F3) and reverse (B3) outer primers. The assay may also include loop forward (LF) and/or loop back (LB) primers to accelerate the reaction. Neisseria meningitides serotypes A, B, C, Y and W135 can be detected using the assay of the invention.

1 . A LAMP assay for detection of meningococcal disease, the test comprising at least one nucleic acid primer set capable of detecting  Neisseria meningitidis  in a LAMP based molecular test, the primer set being chosen from the primer sets listed in Table 1 as LAMP SETS 1 to 12 comprising SEQUENCE IDs from ID: 1 to ID: 69. 
     
     
         2 . An assay as claimed in  claim 1  wherein the primer set consists of one pair of forward (FIP) and reverse (BIP) inner primers, forward (F3) and reverse (B3) outer primers. 
     
     
         3 . An assay as claimed in  claim 1  further comprising loop forward (LF) and/or loop back (LB) primers. 
     
     
         4 . A LAMP assay as claimed in  claim 1  for detection of  Neisseria meningitidis , the assay using primer LAMP SET L3L3 consisting of Seq IDs 11 to 14, 16 and 17. 
     
     
         5 . A LAMP assay as claimed in  claim 1  for detection of  Neisseria meningitidis , the assay using primer LAMP SET 3 L3L1 consisting of Seq IDs 11 to 16. 
     
     
         6 . A LAMP assay as claimed in  claim 1  for detection of  Neisseria meningitidis , the assay using LAMP primer SET 5 L5L1 consisting of Seq IDs 24 to 29. 
     
     
         7 . A method for preparing an assay for the detection of  Neisseria meningitidis  comprising utilizing a primer as set out in Table 1 as Sequence ID: 1 through Sequence ID: 69, individually or in combination with any of the other listed primers in the preparation of the assay. 
     
     
         8 . Use of the primers as claimed in  claim 1  wherein the assay is a LAMP assay. 
     
     
         9 . A method for the detection of  Neisseria meningitidis  serotypes A, B, C, Y or W 135 from a sample comprising a LAMP assay for detection of  Neisseria meningitidis  serotypes A, B, C, Y or W 135, the test comprising at least one nucleic acid primer set capable of detecting  Neisseria meningitidis  in a LAMP based molecular test, the primer set being chosen from the primer sets listed in Table 1 as LAMP SETS 1 to 12 comprising SEQUENCE IDs from ID: 1 to ID: 69. 
     
     
         10 . A method of diagnosing meningococcal disease in a patient, the method comprising the steps of taking a sample from the patient, accessing DNA in the sample, or extracting DNA from the sample, using at least one LAMP primer set as set out on Table 1 consisting of primer sequences chosen from sequence IDs 1 to 69 to amplify the DNA and testing for amplification to confirm presence of  Neisseria meningitidis.