What is the 14-Pathogen Febrile Syndrome PCR Panel (incl. Borrelia burgdorferi, Pneumocystis, Aspergillus)

The iCassette – 14-Pathogen Febrile Syndrome PCR Panel is a multiplex molecular diagnostic solution designed to identify key pathogens associated with fever of unknown origin (FUO). The panel detects 14 targets in a single closed-tube reaction, reducing sample handling and potential contamination. Among the targets are clinically relevant bacteria, fungi, and zoonotic agents — including Borrelia burgdorferi, Pneumocystis jirovecii, Aspergillus, and Cryptococcus.

Each pathogen is identified by a unique melting temperature (Tm) signature using melting curve analysis, ensuring high-resolution discrimination even in polymicrobial samples. The assay is optimized for oropharyngeal swabs, bronchoalveolar lavage fluid, stool, and cerebrospinal fluid (CSF), providing flexibility across clinical presentations.

Used in combination with the Galaxy instrument series (Neo, Lite, Pro), this panel allows for rapid, fully automated detection with minimal manual steps. Laboratories can implement broad-spectrum febrile pathogen detection within standard workflows, supported by internal and external quality controls for high confidence in results.

Role of Pneumocystis and Other Targets in Disease

The pathogens included in this panel represent a wide clinical spectrum, ranging from community-acquired infections to opportunistic and travel-related diseases. For immunocompromised patients, Pneumocystis jirovecii is a major concern. It causes Pneumocystis pneumonia (PCP), a life-threatening infection that often presents with persistent fever, cough, and hypoxia. Early detection is critical for guiding antifungal therapy.

Borrelia burgdorferi, the causative agent of Lyme disease, may lead to recurrent fever, neurological symptoms, or arthritic complications. Other bacterial pathogens like Salmonella typhi, Yersinia pestis, and Neisseria meningitidis are associated with systemic infections, some of which require immediate intervention.

Fungal pathogens such as Aspergillus and Cryptococcus are also included in the panel, both of which are known to cause pulmonary or CNS infections, particularly in immunocompromised individuals. Zoonotic organisms like Brucella, Leptospira, Ehrlichia, and Anaplasma are often underdiagnosed but play a significant role in febrile illness globally.

By including these diverse organisms — with Pneumocystis as one of the critical fungal markers — the panel offers an advanced diagnostic platform for complex febrile syndromes.

About the iCassette – 14-Pathogen Febrile Syndrome Detection Kit

This diagnostic kit is based on multiplex PCR with melting curve analysis, integrated into a proprietary iCassette format. The iCassette is a pre-filled, single-use cartridge containing all necessary reagents for nucleic acid extraction, amplification, and detection.

Pathogens detected:

• Borrelia burgdorferi

• Pneumocystis jirovecii

• Aspergillus spp.

• Cryptococcus spp.

• Neisseria meningitidis

• Salmonella typhi

• Salmonella paratyphi

• Yersinia pestis

• Streptococcus suis

• Group A Streptococcus

• Brucella spp.

• Leptospira spp.

• Anaplasma spp.

• Ehrlichia spp.

The melting curve approach enables simultaneous detection of all targets in one reaction tube. Each organism is identified by a distinct Tm value, eliminating the need for multiple assays. This makes it especially effective for identifying co-infections, such as Pneumocystis in patients with concurrent bacterial sepsis or viral infections.

The kit includes 12 iCassettes per box, positive and negative control reagents, lyophilized PCR reagents, and sterile deionized water. The reagents are batch-specific and must be used with compatible instruments.

Principle and Workflow

The kit’s workflow is fully automated when used with Galaxy Neo, Lite, or Pro qPCR systems. It simplifies the diagnostic process with minimal human intervention:

1. Sample Preparation
   Depending on the sample type, basic pretreatment (vortexing, centrifugation, dilution) may be required. For BAL or CSF, samples are typically ready for direct input.

2. Cassette Loading
   The user adds 1 mL of processed sample to the iCassette sample well, closes the lid, and inserts the cassette into the instrument.

3. Automated Execution
   The system performs nucleic acid extraction, amplification, and melting curve detection in a sealed environment. The QR code on the cassette ensures the correct protocol is loaded.

4. Data Interpretation
   Upon completion, the software outputs the Tm values for each detected pathogen. Pneumocystis is detected within the 60.0–64.0 °C Tm range, with high specificity.

5. Quality Control
   Internal control (RNase P) and pathogen-specific controls validate every run. Invalid runs are automatically flagged.

This seamless process enables high-throughput diagnostics without compromising result integrity or traceability.

Clinical Application

This panel is ideally suited for hospital labs, infectious disease departments, and reference centers dealing with:

• Fever of unknown origin (FUO)

• Travel-related febrile illnesses

• Opportunistic infections in immunocompromised patients

• CNS infections with unclear etiology

• Sepsis and atypical pneumonia diagnostics

The detection of Pneumocystis alongside bacterial and other fungal pathogens enables better clinical decision-making, particularly in complex cases where coinfections are possible. Rapid turnaround and broad-spectrum coverage allow clinicians to initiate targeted therapy early, improving outcomes and supporting antimicrobial stewardship.

The panel also reduces the need for multiple sample draws and separate tests, streamlining diagnostics for both patient and laboratory.