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. 2020 Sep 23:10:528884.
doi: 10.3389/fcimb.2020.528884. eCollection 2020.

Leishmaniasis Diagnosis via Metagenomic Next-Generation Sequencing

Hongbin Chen  1 , Chunhong Fan  1 , Hua Gao  1 , Yuyao Yin  1 , Xiaojuan Wang  1 , Yawei Zhang  1 , Hui Wang  1
Affiliations

Leishmaniasis Diagnosis via Metagenomic Next-Generation Sequencing

Hongbin Chen et al. Front Cell Infect Microbiol. .

Abstract

Leishmaniasis is a vector-borne disease caused by Leishmania. Although the incidence of leishmaniasis in China is currently low, it has not been completely eradicated. In 2019, visceral leishmaniasis was diagnosed in three patients using bone marrow microscopic examination and metagenomic next-generation sequencing (mNGS). The bone marrow mNGS results from the three patients indicated that 99.9, 99.6, and 30.3% of non-human reads matched the Leishmania genome, and plasma mNGS results from one of the patients revealed that 46.2% of non-human reads matched the Leishmania genome. In the second patient's plasma, no Leishmania sequences were detected by plasma mNGS, and the third patient's plasma was unavailable. The pathogen in all three patients was identified as Leishmania infantum. Leishmania amastigotes were observed by microscopic examination of bone marrow smears in all three patients, but were not found in peripheral blood smears. This indicates that the sensitivity of mNGS is higher than that of smear microscopy and that mNGS can be used to identify Leishmania at the species level. All three patients were elderly male farmers, two from Shanxi and one from Beijing. All three patients had splenomegaly and pancytopenia. Originally, these patients were misdiagnosed and treated for extended periods in other hospitals. Diagnoses of visceral leishmaniasis took place 6, 2, and 2 months after the onset of symptoms in the three patients. In conclusion, this study confirms that bone marrow mNGS can be used to quickly and accurately confirm a diagnosis in patients with suspected leishmaniasis.

Keywords: Leishmania; diagnosis; infection; leishmaniasis; mNGS.

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Figures

Figure 1
Figure 1
(A) Arrowheads show Leishmania amastigotes, which are oval and 2.9–5.7 × 1.8–4.0 μm in size. The cytoplasm is stained lilac or purple-blue by Wright's stain and contains a large round nucleus. The nucleus (red or lavender) is located at the front of the worm and accounts for a third to a half of the worm's length. The moving matrix is located next to the nucleus and is rod-shaped and dark in color. (B) The distribution of non-human sequences identified in patients' bone marrow.
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