Compositions and methods for diagnosing parasitic infections
Provided herein are compositions and methods for diagnosing parasitic infections. In particular, provided herein are compositions and methods for preparing samples (e.g., canine fecal samples) and performing Raman spectroscopy detection of parasites infections in the samples.
1. A method of identifying the presence of a parasite in a fecal sample from a subject, comprising:
a) diluting said sample to generate a diluted sample, wherein said sample comprises less than 1 gram of feces; and
b) obtaining a Raman spectrum of said sample using a Raman spectrometer, wherein said Raman spectrum identifies the presence of a parasite in said sample, wherein said presence of a parasite in said sample is determined based on the presence of parasite specific coproantigens in said sample.
2. The method of claim 1 , wherein said feces sample comprises 100-200 mg feces.
3. The method of claim 2 , wherein said feces sample comprises 100 mg feces.
4. The method of claim 1 , wherein said Raman spectroscopy utilizes internal reflective amplification.
5. The method of claim 1 , wherein said Raman spectroscopy generates a molecular fingerprint of said sample.
6. The method of claim 5 , wherein said molecular fingerprint comprises spectral bands indicative of said coprantigens.
7. The method of claim 5 , wherein said molecular fingerprint identifies the genus, species, or strain of parasite in said sample.
8. The method of claim 7 , wherein said parasite is selected from the group consisting of hookworm, roundworm, Cystoisospora, Giardia, and whipworm.
9. The method of claim 1 , wherein said Raman spectrum are analyzed using a machine learning algorithm.
10. The method of claim 9 , wherein said machine learning algorithm is support vector machine and/or random forest algorithms.
11. The method of claim 1 , wherein said Raman spectrum are pre-processed prior to said analysis using a machine learning algorithm.
12. The method of claim 11 , wherein said pre-processing comprising one or more of morphology-split background subtraction and unit vector normalization.
13. The method of claim 1 , wherein said method further comprises generating a receiver operating characteristic curve (ROC) for multiple classes.
14. The method of claim 13 , wherein said ROC comprises a micro-averaging step.
15. A method of treating a parasitic infection in a canine subject, comprising:
a) detecting the presence of a parasitic infection in a sample from said canine subject using a method of claim 1 ; and
b) administering an antibiotic to said canine subject.
16. A system or kit comprising:
a) a diluted fecal mixture from a subject, wherein said diluted fecal mixture comprises a fecal sample and a liquid comprising water, wherein said fecal sample comprises less than one gram of feces;
b) a Raman spectrometer; and
c) a computer system comprising a computer memory and computer processor, wherein said computer memory comprises a library of Raman spectrometric signatures from antigens from parasites, and algorithm for comparing a Raman spectrometric signature from said sample to said library, wherein said computer system is operably linked to said Raman spectrometer.