Semiautomation of Nucleic Acid-based Assays for Chlamydia trachomatis and Neisseria gonorrhoeae

Johnson, Shirley J.; Green, Diane H.; Reed, David; Wood, Linda

doi:10.1093/clinchem/47.4.760

Cited by 1 publication

(2 citation statements)

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“…Four target sequences were selected (Table 1) as representatives of two different, broad microbial groups found in environmental samples such as seawater (Escherichia coli and Candida albicans) 17 or of two pathogens that may occur individually or simultaneously in human urogenital swab or urine specimens (Chlamydia trachomatis and Neisseria gonorrhoeae). 18 In order to mimic more closely the natural nucleic acid materials, the synthetic target oligonucleotide sequences exceeded the length of the part (central in each target in Table 1) for which complementary sequences in probes would be constructed. These targets were synthesized on automated oligonucleotide synthesizers by established methods using phosphoramidite monomers, then purified and quantified prior to use.…”

Section: ' Results and Discussionmentioning

confidence: 99%

“…EcoB1932-1947(-)HICS16 1.3 [16] 2.57 ( 0.14 [18] 0.157 ( 0.004 [16] (3.93 ( 0.22) Â 10 7 [16] 4.60 ( 0.33 [16] EcoB1932-1947(-)HICS17 0.63 [16] 4.53 ( 0.15 [18] 0.339 ( 0.019 [16] (2.08 ( 0.06) Â 10 8 [16] 150 ( 6 [16] EcoB1932-1947(-)HICS18 0.70 [6] 3.99 ( 0.07 [6] 0.291 ( 0.008 [14] (3.34 ( 0.09) Â 10 8 [14] 285 ( 10 [14] CalA1185-1206(-)HICS86 1.0 [18] 3.49 ( 0.17 [20] 0.141 ( 0.003 [18] (3.74 ( 0.08) Â 10 8 [18] 73.3 ( 2.8 [18] CalA1185-1206(-)HICS87 1.8 [16] 2.92 ( 0.27 [18] 0.0188 ( 0.0004 [16] (2.27 ( 0.04)…”

Section: ' Results and Discussionmentioning

confidence: 99%

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Simultaneous Quantification of Multiple Nucleic Acid Targets Using Chemiluminescent Probes

Browne¹,

Deheyn

El‐Hiti

et al. 2011

J. Am. Chem. Soc.

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A novel method is described for simultaneous detection and quantification of attomoles or a few femtomoles of two (or potentially more) nucleic acid targets, without need for amplification. The technique depends on spectral-temporal resolution of chemiluminescence emitted from independent hybridization-induced chemiluminescent signal probes. The probes are internally quenched except in the presence of their specific targets, thereby allowing detection limits up to 10,000 times lower than with fluorescent probes. This is sufficient to obviate the need for amplification in many cases. The utility of the technique has been demonstrated by use of resolvable N-linked acridinium and 2,7-dimethoxyacridinium ester labeled probes in a homogeneous assay for sensitive and simultaneous independent quantification of pan-bacterial and pan-fungal target sequences in seawater.

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Section: ' Results and Discussionmentioning

confidence: 99%