Efficiency evaluation of three assays for detection of Schistosoma japonicum infections in wild mice

doi:10.12140/j.issn.1000-7423.2025.02.006

Abstract

Abstract:

Objective To evaluate the performance of liver homogenate smear microscopy (parasitological detection), real-time quantitative reverse transcription PCR (qPCR) assay and loop-mediated isothermal amplification (LAMP) for detection of Schistosoma japonicum infections in wild rodents, so as to provide an optimal laboratory diagnostic assay for surveillance of S. japonicum infection in wild rodents. Methods A total of 115 wild rodents liver samples (63 samples from Village A and 52 samples from Village B) were collected from Dongzhi County, Anhui Province, and detected by liver homogenate smear microscopy, qPCR assay and LAMP, respectively. The positive rates of the samples were calculated and compared. The consistency between qPCR and LAMP for detection of the liver samples was evaluated with microscopy as a standard, and 70% of liver samples with a low consistency between the parasitological assay and nucleic acid tests were randomly selected for validation with DNA sequencing. Results Liver homogenate smear microscopy detected 54 positive liver samples, with a positive rate of 46.96% (54/115), and the positive rate of liver samples was 46.03% (29/63) in Village A and 48.08% (25/52) in Village B (χ² = 0.001, P > 0.05). qPCR assay tested a 69.57% (80/115) positive rate of liver samples, which was higher than liver homogenate smear microscopy (χ² = 11.175, P < 0.01), and there positive rate was 66.67% (42/63) in Village A and 73.08% (38/52) in Village B (χ² = 0.340, P > 0.05). LAMP tested a 56.52% (65/115) positive rate of liver samples, which was comparable to liver homogenate smear microscopy (χ² = 1.741, P > 0.05), and the positive rate was higher in Village A(71.43%, 45/63) than in Village B (38.46%, 20/52) (χ² = 11.293, P < 0.01). There was a high consistency between qPCR assay and liver homogenate smear microscopy (Kappa value = 0.524 4), and a low consistency between LAMP and liver homogenate smear microscopy (Kappa value = 0.154 5). Among the 24 liver samples from Village A that were negative for liver homogenate smear microscopy but positive for LAMP, 17 samples were randomly selected for DNA sequencing, and 15 of these samples showed a sequence identity of 98% and higher with the 28S ribosomal DNA of S. japonicum (GenBank accession No.: JF721395.1) and were therefore identified as positives. Conclusion Three assays have diverse sensitivities for detection of S. japonicum infections in wild rodents. qPCR assay has the highest positive rate for detection of S. japonicum infections in wild rodents and a high consistency with liver homogenate smear microscopy, while LAMP is convenient to perform but has a low consistency with liver homogenate smear microscopy.

Key words: Schistosoma japoniucm, Wild rodents, Microscopic examination of liver homogenates, Real-time quantitative reverse transcription PCR, Loop mediated isothermal amplification

CLC Number:

R532.21

TANG Qi, LV Chao, WANG Xi, GUO Suying, XU Xiaojuan, ZHU Hai, LI Yinlong, LIN Weina, ZHOU Xinjie, FENG Ting, XU Jing, QIN Zhiqiang. Efficiency evaluation of three assays for detection of Schistosoma japonicum infections in wild mice[J]. CHINESE JOURNAL OF PARASITOLOGY AND PARASITIC DISEASES, 2025, 43(2): 186-191.

Figures/Tables 5

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引物名称 Primer name	引物序列 (5′→3′) Primer sequence (5'→3')	扩增片段长度/bp Length of amplified fragment/bp
PCR		405
Sj28_Forward	GGTTTGACTATTATTGTTGAGC
Sj28_Reverse	TCTCACCTTAGTTCGGACTGA
qPCR		165
SjR2_Forward	TGCCAGAGCGATGACCATAAAC
SjR2_Reverse	GCAATGCCTCGCAGTGTGATAG
LAMP		217
Sj28s-F3	GCTTTGTCCTTCGGGCATTA
Sj28s-B3	GGTTTCGTAACGCCCAATGA
Sj28s-FIP	ACGCAACTGCCAACGTGACATACTGGTCGGCTTGTTACTAGC
Sj28s-BIP	TGGTAGACGATCCACCTGACCCCTCGCGCACATGTTAAACTC