# Highly specific and super-sensitive Dot-ELISA and colloidal gold immunochromatographic strips for the detection of Burkholderia glumae and Burkholderia plantarii of Rice bacterial panicle blight

## TL;DR

This paper introduces new, highly sensitive and specific tests for detecting two bacteria that cause rice bacterial panicle blight, enabling rapid and on-site identification.

## Contribution

Development of ultra-sensitive monoclonal antibodies and detection methods (Dot-ELISA and CGICS) for Burkholderia glumae and Burkholderia plantarii.

## Key findings

- Monoclonal antibodies 4A7, 8C5, 12B5, and 14B3 showed high specificity and ultra-sensitivity for B. glumae and B. plantarii.
- Dot-ELISA and CGICS assays detected bacteria at concentrations as low as 9.78 × 10³ CFU/mL.
- The new methods are 2–8 times more sensitive than conventional PCR.

## Abstract

Burkholderia glumae (B. glumae) and Burkholderia plantarii (B. plantarii) are primary causal agents of rice bacterial panicle blight (RBPB) and cause substantial yield losses in rice worldwide. Given their seed-borne transmission characteristics, quarantine status and destructive hazards, rapid, super-sensitive, highly specific on-site detection technologies are urgently needed. Here, using B. glumae Os48 and B. plantarii ZJ171 as immunogens, we prepared two highly specific and ultra-sensitive monoclonal antibodies (mAbs) against B. glumae (4A7 and 8C5) and two highly specific and ultra-sensitive mAbs against B. plantarii (12B5 and 14B3). We then developed dot-enzyme-linked immunosorbent assays (Dot-ELISA) and colloidal gold immunochromatographic strip (CGICS) assays for detecting B. glumae and B. plantarii with the prepared mAbs as the detection antibodies. These developed mAb-based serological techniques enabled the rapid, broad-spectrum, and specific detection of B. glumae and B. plantarii, respectively, and showed no cross-reaction with other control plant bacteria included in the analysis. Moreover, the detection limits of the Dot-ELISAs for B. glumae or B. plantarii were up to 1.96 × 104 colony-forming units (CFU)/mL, and CGICSs could detect B. glumae and B. plantarii at concentrations as low as 9.78 × 103 CFU/mL. Surprisingly, these two serological techniques are 2 ~ 8 times more sensitive than conventional polymerase chain reaction (PCR). Collectively, the two newly developed serological techniques in this work provide two simple, rapid, broad-spectrum, highly specific, ultra-sensitive and on-site means to detect B. glumae and B. plantarii in rice grains and leaves, thereby offering reliable and practical tools for the detection and quarantine of these two RBPB pathogens.

The online version contains supplementary material available at 10.1007/s44297-026-00067-6.