Method and system for rapid malaria detection

• US 9,983,130 B2
• Filed: 02/05/2014
• Issued: 05/29/2018
• Est. Priority Date: 02/05/2014
• Status: Active Grant

• Current Assignee: Monash University, University of Melbourne
• Original Assignee: Monash University, University of Melbourne
• Inventors: Wood, Bayden Robert, Khoshmanesh, Aazam, Dixon, Matthew, Tilley, Leann, McNaughton, Donald
• Primary Examiner(s): Kim, Kiho
• Application Number:
  US15/117,149
• Publication Number:
  US 20170167976A1
• Time in Patent Office: 1,574 Days
• CPC Class Codes: G01N 2021/3595 using FTIR
• CPC Class Codes: G01N 21/3577 for analysing liquids, e.g. polluted water
• CPC Class Codes: G01N 21/552 Attenuated total reflection
• CPC Class Codes: G01N 2201/129 Using chemometrical methods
• CPC Class Codes: G01N 33/49 Blood chemical methods for determining blood cell populations G01N33/5094; chemical analysis of blood groups or blood types G01N33/80

Abstract

The invention described herein relates to a method of detecting malaria comprising the steps of: (i) delivering an evanescent IR beam through said ATR substrate in contact with a patient blood sample; (ii) detecting IR radiation transmitted from the ATR substrate to produce a signal characteristic for one or more lipids in the sample, and (iii) processing said signal and a set of reference library spectra of lipids associated with malaria parasites in order to detect matches and quantify said one or more lipids in the sample. In contrast to the prior art, the present invention relies on detecting lipids instead of hemozoin.

Claims

1. A method of detecting malaria with an ATR-FTIR spectrometer comprising:

(i) delivering an evanescent IR beam using an ATR substrate of an ATR-FTIR spectrometer in contact with a patient sample, wherein the patient sample comprises red blood cells at least partially concentrated and separated from a patient blood sample and dried onto the ATR substrate;

(ii) detecting IR radiation transmitted from the ATR substrate to produce a spectrum of the patient sample;

(iii) processing one or more signals in the spectrum to identify any malaria parasite; and

(iv) determining a parasitemia level of the patient sample using one or more signals in the spectrum.

2. The method according to claim 1 further comprising detecting a ring form or a gametocyte form of malaria parasite.

3. The method according to claim 1 wherein the parasitemia level is ≥

100 parasites/μ

l of sample.

4. The method according to claim 1 wherein the parasitemia level is ≥

50 parasites/μ

l of sample.

5. The method according to claim 1 wherein the processing comprises converting the spectrum into a second derivative, then applying a partial least squares regression model generated by using a library comprising a calibration set of spectral standards containing mixtures of normal and infected blood at different ratios.

6. The method of claim 1, further comprising fixing the sample in methanol.

7. The method of claim 6, further comprising drying the sample using a heat source.

8. The method of claim 1, wherein the one or more signals in the spectra are one more signals in the spectra at 2850, 2854, 2873, 2904, 2908, 2942, 2978, 2982, 2985, 3012 or 3040 cm⁻

1.

9. The method of claim 1, wherein the one or more signals in the spectrum comprise different spectra at different parasitemia levels.

10. The method of claim 1, wherein the one or more signals in the spectrum comprise at least one signal at 2854, 2904, 2942 or 2982 cm⁻

1 to quantify a parasitemia level between 10-30%.

11. The method of claim 1, wherein the one or more signals in the spectrum comprise signals at 2854, 2904, 2942 and 2982 cm⁻

1 to quantify a parasitemia level between 10-30%.

12. The method of claim 1, wherein the one or more signals in the spectra comprise at least one signal of at 2873, 2908, 2978 or 3012 cm⁻

1 to quantify a parasitemia level between 0-5%.

13. The method of claim 1, wherein the one or more signals in the spectrum comprise signals at 2873, 2908, 2978 and 3012 cm⁻

1 to quantify a parasitemia level between 0-5%.

14. The method of claim 1, wherein the one or more signals in the spectrum comprise at least one signal of at 2857, 2942, 2985 or 3040 cm⁻

1 to quantify a parasitemia level between 0-1%.

15. The method of claim 1, wherein the one or more signals in the spectra comprise signals at 2857, 2942, 2985 and 3040 cm⁻

1 to detect the presence of any parasitemia.

17. The method of claim 1, wherein the processing of the signal is performed via execution by the processor of a predetermined instruction set stored in a computer readable medium.

16. A system of analysis for diagnosing malaria, the system comprising:

an ATR substrate for receiving a blood sample;

an FTIR spectrometer configured to generate an evanescent wave that extends into the sample using the ATR substrate;

a detector for detecting IR radiation transmitted from the ATR substrate to produce a signal; and

a processor configured to process the signal to create an FTIR spectrum of the blood sample and determine a parasitemia level using different wavenumber values for different parasitemia levels.

18. A method, comprising:

with a processor, executing an instruction set stored in a non-transitory computer readable storage medium, the instruction set including instructions to compare one or more spectral signals characteristic of a patient blood sample infected with malaria parasites to detect matches and quantify a parasitemia level using different wavenumber values or absorbance values for different parasitemia levels, associated with at least one parasite chosen from the group comprising P. falciparum, P. vivax, P. ovale, P. malariae and P. knowlesi.

19. The method of claim 18, wherein the one or more signals are spectra at 2850, 2854, 2873, 2904, 2908, 2942, 2978, 2982, 2985, 3012 or 3040 cm⁻

1.

20. The method of claim 18, wherein the one or more signals comprise different spectra at different parasitemia levels.