RAPID PERTUSSIS DIAGNOSIS ON A POINT-OF-CARE HYBRID MICROFLUIDIC BIOCHIP
Certain embodiments are directed to a point-of-care (POC) microfluidic biochip for rapid, highly sensitive and specific pertussis diagnosis. The POC biochip can be used in various venues such as physician's office, schools, hospitals, and low-resource settings so that rapid prevention and treatment of pertussis can be achieved. The POC biochip based pertussis diagnosis is low-cost and does not rely on any specialized instrument, but offers comparable sensitivity to real-time PCR.
1 . A battery-powered loop-mediated isothermal amplification (LAMP) system comprising:
a microfluidic device comprising at least one amplification zone containing amplification primers that differentially amplify Bordetella pertussis ( B. pertussis ) DNA;
a battery operated heater configured to maintain the amplification zone at an amplification temperature; and
a simple detection system.
2 . The system of claim 1 , wherein qualitative results for B. pertussis detection can be visible to the naked eye under a portable UV light source or recorded by a smartphone camera, without the use of specialized instruments.
3 . The system of claim 1 , wherein detection results can be quantified by a low-cost portable battery-powered spectrometer.
4 . The system of claim 1 , wherein the microfluidic device comprises a positive control and a negative control for B. pertussis DNA amplification.
5 . The system of claim 1 , wherein the microfluidic device is composed of paper and polymers as the device substrate.
6 . The system of claim 5 , wherein paper is used to preload primers, enabling longer shelf life.
7 . The system of claim 1 , wherein the microfluidic device is configured to receive two or more samples.
8 . The system of claim 1 , further comprising a user interface.
9 . The system of claim 1 , wherein the battery is a 9-volt battery.
10 . A method of detecting Bordetella pertussis in a patient sample comprising:
performing bacterial lysis on a sample suspected of having B. pertussis bacteria;
introducing the lysed sample suspected of having B. pertussis bacteria into a microfluidic device configured to specifically amplify B. pertussis DNA;
incubating the microfluidic device at a temperature for LAMP amplification of B. pertussis DNA; and
detecting the presence or absence of B. pertussis DNA based on DNA amplification.
11 . The method of claim 10 , wherein the LAMP amplification temperature is 55° C. to 65° C.
12 . The method of claim 10 , wherein the microfluidic device is configured with a positive control and a negative control for B. pertussis DNA amplification.
13 . The method of claim 10 , further comprising exposing the sample to a bacterial lysis solution, without centrifuges or other equipment for cell lysis.
14 . The method of claim 10 , wherein the sensitivity and specificity for clinical sample testing are 100% and 96%, respectively.