First Case Series of Emerging Rickettsial Neonatal Sepsis Identified by Pcr Based Dna Sequencing
Title of the paper: First case series of emerging Rickettsial neonatal sepsis identified by PCR based DNA sequencing
ABSTRACT
Purpose : To detect and identify the etiological agent in the peripheral blood from the cases of neonatal sepsis.
Methods: Four neonates from geographically different regions of South India presented with signs of neonatal sepsis and all the routine clinical and laboratory investigations were performed. Blood culture by Bac T Alert 3D, was negative. To establish the aetiology, Polymerase Chain reaction (PCR) for eubacterial genome and subsequent amplification with Gram positive and Gram negative primers were performed followed by DNA sequencing.
Results : PCR for the detection of eubacterial genome was positive in all the four neonates and further amplification with designed Gram positive and Gram negative primers revealed the presence of Gram negative bacteria. The amplicons were identified as Orientia tsutsugamushi in three neonates and Coxiella burnetti in the other neonate. Multalin analysis was done to further characterize the strain variation among the three strains.
Conclusion : PCR based DNA sequencing is a rapid and reliable diagnostic tool to identify the aetiological agents of neonatal sepsis. This is the first case series of emerging Rickettsial neonatal sepsis in India .
Number of words: 181
Key words : Orientia tsutsugamushi, Coxiella burnetti neonatal sepsis,
doxycycline, PCR based DNA sequencing
Introduction
Rickettsial infections are re-emerging and are prevalent throughout the world. In India, they are reported from Maharashtra, Tamil nadu, Karnataka, Kerala, Jammu and Kashmir, Uttaranchal, Himachal Pradesh, Rajasthan, Assam and West Bengal. In view of low index of suspicion, nonspecific signs and symptoms, and absence of widely available sensitive and specific diagnostic test, these infections are
ABSTRACT
Purpose : To detect and identify the etiological agent in the peripheral blood from the cases of neonatal sepsis.
Methods: Four neonates from geographically different regions of South India presented with signs of neonatal sepsis and all the routine clinical and laboratory investigations were performed. Blood culture by Bac T Alert 3D, was negative. To establish the aetiology, Polymerase Chain reaction (PCR) for eubacterial genome and subsequent amplification with Gram positive and Gram negative primers were performed followed by DNA sequencing.
Results : PCR for the detection of eubacterial genome was positive in all the four neonates and further amplification with designed Gram positive and Gram negative primers revealed the presence of Gram negative bacteria. The amplicons were identified as Orientia tsutsugamushi in three neonates and Coxiella burnetti in the other neonate. Multalin analysis was done to further characterize the strain variation among the three strains.
Conclusion : PCR based DNA sequencing is a rapid and reliable diagnostic tool to identify the aetiological agents of neonatal sepsis. This is the first case series of emerging Rickettsial neonatal sepsis in India .
Number of words: 181
Key words : Orientia tsutsugamushi, Coxiella burnetti neonatal sepsis,
doxycycline, PCR based DNA sequencing
Introduction
Rickettsial infections are re-emerging and are prevalent throughout the world. In India, they are reported from Maharashtra, Tamil nadu, Karnataka, Kerala, Jammu and Kashmir, Uttaranchal, Himachal Pradesh, Rajasthan, Assam and West Bengal. In view of low index of suspicion, nonspecific signs and symptoms, and absence of widely available sensitive and specific diagnostic test, these infections are
References: an international perspective. Arch Dis Child Fetal Neonatal Ed. 2005; 90: 220 – 24.