Table 2.

Comparison of existing parasitological detection methods against our method

	Parasite	T. cruzi		T. brucei gambiense		T. brucei rhodesiense		Blood	CSF	LoD/Sensitivity/Specificity	Blood volume analyzed	CSF volume analyzed	Expert requirement (e.g., benchtop microscope)	Benchtop device requirement for sample preparation (e.g., centrifugation)	Electricity requirement	Field-portable	Microscopic examination of parasites manually	Total analysis time per test
	Stage	Acute	Chronic	I	II	I	II
mAECT tubesa 25				✓	✓			✓		<50 trypanosomes/mL	350 μL/test		Yes	Yes	Yes	No	Yes	45–60 min
PCR34		✓	✓	✓34	✓34	✓35	✓34	✓	✓	High sensitivity and specificity for T. brucei gambiense and T. cruzi36 Low sensitivity for T. brucei rhodesiense35	200 μL34	1000 μL34	Yes	Yes	Yes	Yes	No	~5 hb
Thick smear37		✓	✓	✓	✓	✓	✓	✓	✓	Low sensitivity	2–4 μL/slide	2–4 μL/slide	Yes	Yes	Yes	No	Yes	75–90 min
Microhematocrit		✓38		✓18	✓18	✓18	✓18	✓18	✓18	High sensitivity for T. b. gambiense and T.b. rhodesiense (an estimated detection threshold of 500 trypanosomes/ml)18	6 × 50–75 μL/tube		Yes	Yes	Yes	No	Yes	~15 min38
Strout method39		✓		✓	✓	✓	✓	✓		Low sensitivity	2–4 μL/slidec		Yes	Yes	Yes	No	Yes	~30 mind
Card agglutination test for trypanosomiasis (CATT)39				✓	✓			✓		High specificity and sensitivity for T.b. gambiensee Not for T. cruzi and T.b. rhodesiense	30–50 μL40		No	No	Nof	Yes	No	5 min
Rapid diagnostic tests4,40		✓41		✓	✓			✓		Low specificity, especially for low parasitemia levels4	30 μL42		No	No	No	Yes	No	15 min42
Our method		✓		✓	✓	✓	✓	✓	✓	10 trypanosomes/mL for blood; 3 trypanosomes/mL for CSF	~800 μL	~3200 μL	No	No	Yes	Yes	—	~20 min

The bold and italics text/values were used to emphasize the row that includes the specifications of our method

^aThis method can be made even more sensitive (10 trypanosomes/mL) by performing centrifugation of whole blood and loading buffy coat onto the mAECT tube⁴³

^bIncluding DNA isolation, PCR, and electrophoresis

^cThe blood analyzed (2–4 μL) is taken from a centrifuged/concentrated blood sample

^d5 min for centrifugation plus time required to analyze each slide

^eCATT does not work for some strains of T. b. gambiense because they lack the specific antigen; and its specificity is low in areas with low prevalence⁴

^fCATT requires a rotator to mix the blood sample for 5 min at 60 rpm