<p>Citrus industry faces significant threats from diverse pathogens, however the two major pathogens, citrus tristeza virus (CTV) and <i>Candidatus</i> Liberibacter asiaticus (<i>C</i>Las), have severely impacted fruit quality, yield and orchards longevity. Diagnosis of disease caused by such devastating pathogens with the rapid and accurate detection method is very crucial for early disease management. Several methods are employed routinely for detection of CTV and CLas, however most of them have limitations such as being labour-intensive procedures, time-consuming, and lacking in sensitivity and specificity, which leads to limits their suitability for routine screening and management. In the present study, we developed a duplex polymerase chain reaction coupled with a lateral flow immunoassay (d-PCR-LFIA) for rapid and simultaneous detection of CTV and <i>C</i>Las pathogens. The developed d-PCR-LFIA method can be exploited for onsite detection of these two major pathogens, since it can eliminate the agarose gel electrophoresis-based end point detection. The developed diagnostic tools offer high sensitivity and ease of interpretation for rapid detection by combining the specificity of PCR with the simplicity of LFIA visualization. The assay was standardized by designing and labelling the specific primer pairs based on the conserved region of the coat protein gene <i>p25</i> of CTV (CTRPA-F/R) and <i>16S rRNA</i> gene of <i>C</i>Las (DKG-F/R). A series of parameters were systematically tested and fine-tuned to achieve optimal assays performance. The optimal limit of detection (LOD) was achieved at ≤0.135 pg for CTV and 40 pg of <i>C</i>Las, cDNA/DNA respectively. Also, the sensitivity of d-PCR-LFIA product was achieved at a LOD of 1x10^-4 µL, after dilution of labelled d-PCR amplicon. The specificity of the assay was determined and validated by using a total 90 samples of known positive, field, and healthy control. The assay did not show any cross reactivity with other citrus pathogens and showed 100 % specificity with CTV and <i>C</i>Las pathogens. Cohen's Kappa value of 0.977 indicates an almost perfect agreement, demonstrating a high level of reliability between the evaluated diagnostics tools. This method enables the rapid, naked-eye based visual detection of CTV and <i>C</i>Las without being contacted to carcinogenic chemicals. The d-PCR-LFIA represents a significant advancement in the field of disease diagnostics, and it has a potential to become a point-of-care diagnostic tools for simultaneous detection of both pathogens (CTV and <i>C</i>Las) and might be useful in mobile laboratories with portable equipment (‘lab in a van’) to conduct citrus clean plant program.</p>