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Development of a loop-mediated isothermal amplification-lateral flow dipstick (LAMP-LFD) assay and on-site rapid detection evaluation for Rodentibacter heylii and Rodentibacter pneumotropicus
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Huiqiong Yan1, 2, 3, Sisi Chen1, 2, Yuhan Gan4, Jing Xing5, Yufang Feng5, Xin Wen1, 2, Jie Fang1, 2, Jiangtao Du1, 2, Shasang Zhou1, 2, Honggang Guo1, 2, Xiaoyin Jin1, 2, Zhiyuan Wang1, 2, Junhao Tao1, 2, Lingqun Lu1, 2, Qingming Kong6, Huazhong Ying1, 2, 7, Wei Han1, 2, 7, 8, Fangwei Dai1, 2, 7, 8
Animal Models and Experimental Medicine | 2026, 9(7) : 1395 - 1408
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Animal Models and Experimental Medicine | 2026, 9(7): 1395-1408
• Regular Articles •
Development of a loop-mediated isothermal amplification-lateral flow dipstick (LAMP-LFD) assay and on-site rapid detection evaluation for Rodentibacter heylii and Rodentibacter pneumotropicus
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Huiqiong Yan1, 2, 3, Sisi Chen1, 2, Yuhan Gan4, Jing Xing5, Yufang Feng5, Xin Wen1, 2, Jie Fang1, 2, Jiangtao Du1, 2, Shasang Zhou1, 2, Honggang Guo1, 2, Xiaoyin Jin1, 2, Zhiyuan Wang1, 2, Junhao Tao1, 2, Lingqun Lu1, 2, Qingming Kong6, Huazhong Ying1, 2, 7, Wei Han1, 2, 7, 8, Fangwei Dai1, 2, 7, 8
Affiliations
  • 1Center of Laboratory Animal, Hangzhou Medical College, Hangzhou, Zhejiang, China
  • 2Zhejiang Provincial Key Laboratory of Laboratory Animals and Safety Research, Hangzhou Medical College, Hangzhou, Zhejiang, China
  • 3Department of Pathology, Cancer Prevention and Treatment Institute of Chengdu, Chengdu Fifth People's Hospital (The Second Clinical Medical College, Affiliated Fifth People's Hospital of Chengdu University of Traditional Chinese Medicine), Chengdu, Sichuan, China
  • 4Savaid Stomatology School, Hangzhou Medical College, Hangzhou, Zhejiang, China
  • 5National Rodent Laboratory Animal Resources Center, Institute for Laboratory Animal Resources, National Institutes for Food and Drug Control (NIFDC), Beijing, China
  • 6Key Laboratory of Biomarkers and In Vitro Diagnosis Translation of Zhejiang Province, School of Laboratory Medicine and Bioengineering, Hangzhou Medical College, Hangzhou, Zhejiang, China
  • 7Engineering Research Center of Novel Vaccine of Zhejiang Province, Hangzhou Medical College, Hangzhou, Zhejiang, China
  • 8Zhejiang Key Laboratory of High-level Biosafety and Biomedical Transformation, Hangzhou Medical College, Hangzhou, China
Published: 2026-07-28 doi: 10.1002/ame2.70214
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Background:

Rodentibacter pneumotropicus and Rodentibacter heylii are important exclusion pathogens in specific pathogen-free (SPF) laboratory animals. However, routine detection methods such as traditional bacterial culture remain labor-intensive and time-consuming. The loop-mediated isothermal amplification-lateral flow dipstick (LAMP-LFD) assay provides a self-contained format for nucleic acid testing that combines speed, sensitivity, and specificity while minimizing aerosol contamination, making it well-suited for point-of-need applications.

Objectives:

This study aimed to develop a streamlined, on-site detection platform capable of rapidly identifying both pathogens with high analytical accuracy, operational simplicity, and suitability for routine laboratory surveillance.

Methods:

A LAMP-LFD system was established for the detection of the two pathogens using primers designed against the conserved 16S-23S rRNA ITS region. The assay's specificity, sensitivity, repeatability, and limit of detection (LoD) were evaluated.

Results:

The assay achieved detection limits of 1 × 10−4 ng/μL for R. pneumotropicus and 1 × 10−5 ng/μL for R. heylii, demonstrating significantly higher sensitivity than PCR. No cross-reactivity was observed with 15 non-target bacteria. Among 28 infected samples, the method detected 19 positives, compared with 21 by multiplex PCR and 23 by qPCR. In 842 laboratory animal samples, culture-based, qPCR, and LAMP-LFD detected 2, 41, and 37 positive samples, respectively. The LoDs were 102 and 101 CFU/mL in fecal samples and 101 and 100 CFU/mL in throat swab samples for R. pneumotropicus and R. heylii, respectively.

Conclusions:

This method provides an approach for monitoring the health status of laboratory mice by enabling the detection of these two target pathogens.

LAMP-LFD  /  novel detection method  /  Rodentibacter heylii  /  Rodentibacter pneumotropicus
Huiqiong Yan, Sisi Chen, Yuhan Gan, Jing Xing, Yufang Feng, Xin Wen, Jie Fang, Jiangtao Du, Shasang Zhou, Honggang Guo, Xiaoyin Jin, Zhiyuan Wang, Junhao Tao, Lingqun Lu, Qingming Kong, Huazhong Ying, Wei Han, Fangwei Dai. Development of a loop-mediated isothermal amplification-lateral flow dipstick (LAMP-LFD) assay and on-site rapid detection evaluation for Rodentibacter heylii and Rodentibacter pneumotropicus[J]. Animal Models and Experimental Medicine, 2026 , 9 (7) : 1395 -1408 . DOI: 10.1002/ame2.70214
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Funding
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  • Medical Science and Technology Project of Zhejiang Province(2018KY351; 2020KY530)
  • Scientific research project of Zhejiang Graduate Education Society(2021009; 2023027)
  • Basic Public Welfare Research Program of Zhejiang Province(LGD22C040024; ZCLTGD24C0401)
  • Zhejiang Traditional Chinese Medicine Administration(2021ZB081)
  • Department of Education of Zhejiang Province(Y201942573)
  • Project of the Central Government leading Local Scientific and Technological Development Fund(2025ZY01054)
  • Project of Zhejiang Key Laboratory(2025E10040)
  • Postgraduate Research Project and construction project of Hangzhou Medical College(26100548300-6; 26100548300-10)
Appendix
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Year 2026 volume 9 Issue 7
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Article Info
doi: 10.1002/ame2.70214
  • Receive Date：2025-12-10
  • Online Date：2026-08-06
  • Published：2026-07-28
Article Data
Affiliations
History
  • Received：2025-12-10
  • Revised：2026-03-13
  • Accepted：2026-04-06
Funding
Medical Science and Technology Project of Zhejiang Province(2018KY351; 2020KY530)
Scientific research project of Zhejiang Graduate Education Society(2021009; 2023027)
Basic Public Welfare Research Program of Zhejiang Province(LGD22C040024; ZCLTGD24C0401)
Zhejiang Traditional Chinese Medicine Administration(2021ZB081)
Department of Education of Zhejiang Province(Y201942573)
Project of the Central Government leading Local Scientific and Technological Development Fund(2025ZY01054)
Project of Zhejiang Key Laboratory(2025E10040)
Postgraduate Research Project and construction project of Hangzhou Medical College(26100548300-6; 26100548300-10)
Affiliations
    1Center of Laboratory Animal, Hangzhou Medical College, Hangzhou, Zhejiang, China
    2Zhejiang Provincial Key Laboratory of Laboratory Animals and Safety Research, Hangzhou Medical College, Hangzhou, Zhejiang, China
    3Department of Pathology, Cancer Prevention and Treatment Institute of Chengdu, Chengdu Fifth People's Hospital (The Second Clinical Medical College, Affiliated Fifth People's Hospital of Chengdu University of Traditional Chinese Medicine), Chengdu, Sichuan, China
    4Savaid Stomatology School, Hangzhou Medical College, Hangzhou, Zhejiang, China
    5National Rodent Laboratory Animal Resources Center, Institute for Laboratory Animal Resources, National Institutes for Food and Drug Control (NIFDC), Beijing, China
    6Key Laboratory of Biomarkers and In Vitro Diagnosis Translation of Zhejiang Province, School of Laboratory Medicine and Bioengineering, Hangzhou Medical College, Hangzhou, Zhejiang, China
    7Engineering Research Center of Novel Vaccine of Zhejiang Province, Hangzhou Medical College, Hangzhou, Zhejiang, China
    8Zhejiang Key Laboratory of High-level Biosafety and Biomedical Transformation, Hangzhou Medical College, Hangzhou, China

Corresponding:

Wei Han, Center of Laboratory Animal, Hangzhou Medical College, Hangzhou 310000, Zhejiang, China. Email:
Fangwei Dai,
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表12种不同金属材料的力学参数
科
Family		 属数
Number of
genus		 种数
Number of
species		 占总种数比例
Percentage of
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Genus		 种数
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鹅膏菌科Amanitaceae 2 11 5.26 鹅膏菌属 Amanita 10 4.78
小菇科 Mycenaceae 2 12 5.74 丝盖伞属 Inocybe 5 2.39
多孔菌科 Polyporaceae 8 14 6.70 蜡蘑属 Laccaria 5 2.39
红菇科 Russulaceae 3 23 11.00 小皮伞属 Marasmius 6 2.87
小菇属 Mycena 11 5.26
光柄菇属 Pluteus 5 2.39
红菇属 Russula 17 8.13
栓菌属 Trametes 5 2.39
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