Article Figures & Data
Figures
- FIG. 1.
Analytical sensitivity of LAMP for detection of the B. pertussis genome. Total DNA from B. pertussis strain Tohama was serially diluted from 1 ng to 1 fg and amplified by LAMP. (A) Real-time turbidity assay with Loopamp real-time turbidimeter. (B) Electrophoretic analysis of LAMP products (2 μl) for a 60-min reaction. Lanes 1 to 7, 1 ng, 100 pg, 10 pg, 1 pg, 100 fg, 10 fg, and 1 fg DNA/tube, respectively; lane N, negative control; lane M, molecular size marker. OD650, optical density at 650 nm.
Tables
- TABLE 1.
Strains used in this study and comparison of LAMP and PCR specificitya
Strain Result byb: LAMPc IS481-PCR PTp1/p2-PCR B. pertussis Tohama + + + Yamaguchi + + + BP256d + + + BP257d + + + BP258d + + + BP259d + + + BP260d + + + BP289d + + + BP290d + + + B. parapertussis ATCC 15237 − − − ATCC 15311 − − − BAA-587 − − − BPP01e − + − B. hinzii ATCC 51730 − − − B. holmesii ATCC 51541 − + − B. avium ATCC 35086 − − − B. bronchiseptica R05f − − − ↵ a For PCR detections, 1 ng of DNA was tested. For LAMP assay, 10 fg DNAs of B. pertussis strains and 1 ng DNAs of other Bordetella species were tested.
↵ b +, LAMP or PCR amplification; −, no LAMP or PCR amplification.
↵ c Turbidity assay with a 60-min reaction.
↵ d Isolated from Japanese patients in the period 2004 to 2005.
↵ e Isolated from a Japanese patient in 2002.
↵ f Isolated from a rabbit in 1995.
- TABLE 2.
LAMP primers for Bordetella pertussis detection
Primer Type Sequence (5′-3′) BP-F3 F3 CCGCATACGTGTTGGCA BP-B3 B3c TGCGTTTTGATGGTGCCT BP-FIP F2-F1c TTGGATTGCAGTAGCGGGATGTGCATGCGTGCAGATTCGTC BP-BIP B1-B2c CGCAAAGTCGCGCGATGGTAACGGATCACACCATGGCA BP-LF LFc ACGGAAGAATCGAGGGTTTTGTAC BP-LB LB GTCACCGTCCGGACCGTG - TABLE 3.
Results of LAMP, IS481-PCR, and PTp1/p2-PCR versus results of culture for clinical specimens
Culture result No. of specimens tested by: LAMPa IS481-PCR PTp1/p2-PCR Positive Negative Total Positive Negative Total Positive Negative Total Positive 5 0 5 5 0 5 4 1 5 Negative 19 88 107 19 88 107 2 105 107 Total 24 88 112 24 88 112 6 106 112 ↵ a Fluorescent-dye-mediated naked-eye visualization with a 40-min reaction.
