Development of an Immunochromatographic Strip Using Conjugated Gold Nanoparticles for the Rapid Detection of <i>Klebsiella pneumoniae</i> Causing Neonatal Sepsis
Neonatal sepsis is a leading cause of death among newborns and infants, especially in the developing world. The problem is compounded by the delays in pinpointing the causative agent of the infection. This is reflected in increasing mortality associated with these cases and the spread of multi-drug-...
| Main Authors: | , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2021-07-01
|
| Series: | Pharmaceutics |
| Subjects: | |
| Online Access: | https://www.mdpi.com/1999-4923/13/8/1141 |
| _version_ | 1827684400065675264 |
|---|---|
| author | Noha M. Elhosseiny Tamer M. Samir Aliaa A. Ali Amani A. El-Kholy Ahmed S. Attia |
| author_facet | Noha M. Elhosseiny Tamer M. Samir Aliaa A. Ali Amani A. El-Kholy Ahmed S. Attia |
| author_sort | Noha M. Elhosseiny |
| collection | DOAJ |
| description | Neonatal sepsis is a leading cause of death among newborns and infants, especially in the developing world. The problem is compounded by the delays in pinpointing the causative agent of the infection. This is reflected in increasing mortality associated with these cases and the spread of multi-drug-resistant bacteria. In this work, we deployed bioinformatics and proteomics analyses to determine a promising target that could be used for the identification of a major neonatal sepsis causative agent, <i>Klebsiella pneumoniae</i>. A 19 amino acid peptide from a hypothetical outer membrane was found to be very specific to the species, well conserved among its strains, surface exposed, and expressed in conditions simulating infection. Antibodies against the selected peptide were conjugated to gold nanoparticles and incorporated into an immunochromatographic strip. The developed strip was able to detect as low as 10<sup>5</sup> CFU/mL of <i>K. pneumoniae</i>. Regarding specificity, it showed negative results with both <i>Escherichia coli</i> and <i>Enterobacter cloacae</i>. More importantly, in a pilot study using neonatal sepsis cases blood specimens, the developed strip selectively gave positive results within 20 min with those infected with <i>K. pneumoniae</i> without prior sample processing. However, it gave negative results in cases infected with other bacterial species. |
| first_indexed | 2024-03-10T08:29:43Z |
| format | Article |
| id | doaj.art-e40f4dd0a6e649ff824aa1087764edd3 |
| institution | Directory Open Access Journal |
| issn | 1999-4923 |
| language | English |
| last_indexed | 2024-03-10T08:29:43Z |
| publishDate | 2021-07-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Pharmaceutics |
| spelling | doaj.art-e40f4dd0a6e649ff824aa1087764edd32023-11-22T09:13:02ZengMDPI AGPharmaceutics1999-49232021-07-01138114110.3390/pharmaceutics13081141Development of an Immunochromatographic Strip Using Conjugated Gold Nanoparticles for the Rapid Detection of <i>Klebsiella pneumoniae</i> Causing Neonatal SepsisNoha M. Elhosseiny0Tamer M. Samir1Aliaa A. Ali2Amani A. El-Kholy3Ahmed S. Attia4Department of Microbiology and Immunology, Faculty of Pharmacy, Cairo University, Cairo 11562, EgyptDepartment of Microbiology and Immunology, College of Pharmaceutical Sciences and Drug Manufacturing, Misr University for Science and Technology, Cairo 12573, EgyptDepartment of Pediatrics, Faculty of Medicine, Cairo University, Cairo 11562, EgyptDepartment of Clinical Pathology, Faculty of Medicine, Cairo University, Cairo 11562, EgyptDepartment of Microbiology and Immunology, Faculty of Pharmacy, Cairo University, Cairo 11562, EgyptNeonatal sepsis is a leading cause of death among newborns and infants, especially in the developing world. The problem is compounded by the delays in pinpointing the causative agent of the infection. This is reflected in increasing mortality associated with these cases and the spread of multi-drug-resistant bacteria. In this work, we deployed bioinformatics and proteomics analyses to determine a promising target that could be used for the identification of a major neonatal sepsis causative agent, <i>Klebsiella pneumoniae</i>. A 19 amino acid peptide from a hypothetical outer membrane was found to be very specific to the species, well conserved among its strains, surface exposed, and expressed in conditions simulating infection. Antibodies against the selected peptide were conjugated to gold nanoparticles and incorporated into an immunochromatographic strip. The developed strip was able to detect as low as 10<sup>5</sup> CFU/mL of <i>K. pneumoniae</i>. Regarding specificity, it showed negative results with both <i>Escherichia coli</i> and <i>Enterobacter cloacae</i>. More importantly, in a pilot study using neonatal sepsis cases blood specimens, the developed strip selectively gave positive results within 20 min with those infected with <i>K. pneumoniae</i> without prior sample processing. However, it gave negative results in cases infected with other bacterial species.https://www.mdpi.com/1999-4923/13/8/1141<i>K. pneumoniae</i>neonatal sepsisimmunochromatographic stripdiagnosisrapidgold nanoparticles |
| spellingShingle | Noha M. Elhosseiny Tamer M. Samir Aliaa A. Ali Amani A. El-Kholy Ahmed S. Attia Development of an Immunochromatographic Strip Using Conjugated Gold Nanoparticles for the Rapid Detection of <i>Klebsiella pneumoniae</i> Causing Neonatal Sepsis Pharmaceutics <i>K. pneumoniae</i> neonatal sepsis immunochromatographic strip diagnosis rapid gold nanoparticles |
| title | Development of an Immunochromatographic Strip Using Conjugated Gold Nanoparticles for the Rapid Detection of <i>Klebsiella pneumoniae</i> Causing Neonatal Sepsis |
| title_full | Development of an Immunochromatographic Strip Using Conjugated Gold Nanoparticles for the Rapid Detection of <i>Klebsiella pneumoniae</i> Causing Neonatal Sepsis |
| title_fullStr | Development of an Immunochromatographic Strip Using Conjugated Gold Nanoparticles for the Rapid Detection of <i>Klebsiella pneumoniae</i> Causing Neonatal Sepsis |
| title_full_unstemmed | Development of an Immunochromatographic Strip Using Conjugated Gold Nanoparticles for the Rapid Detection of <i>Klebsiella pneumoniae</i> Causing Neonatal Sepsis |
| title_short | Development of an Immunochromatographic Strip Using Conjugated Gold Nanoparticles for the Rapid Detection of <i>Klebsiella pneumoniae</i> Causing Neonatal Sepsis |
| title_sort | development of an immunochromatographic strip using conjugated gold nanoparticles for the rapid detection of i klebsiella pneumoniae i causing neonatal sepsis |
| topic | <i>K. pneumoniae</i> neonatal sepsis immunochromatographic strip diagnosis rapid gold nanoparticles |
| url | https://www.mdpi.com/1999-4923/13/8/1141 |
| work_keys_str_mv | AT nohamelhosseiny developmentofanimmunochromatographicstripusingconjugatedgoldnanoparticlesfortherapiddetectionofiklebsiellapneumoniaeicausingneonatalsepsis AT tamermsamir developmentofanimmunochromatographicstripusingconjugatedgoldnanoparticlesfortherapiddetectionofiklebsiellapneumoniaeicausingneonatalsepsis AT aliaaaali developmentofanimmunochromatographicstripusingconjugatedgoldnanoparticlesfortherapiddetectionofiklebsiellapneumoniaeicausingneonatalsepsis AT amaniaelkholy developmentofanimmunochromatographicstripusingconjugatedgoldnanoparticlesfortherapiddetectionofiklebsiellapneumoniaeicausingneonatalsepsis AT ahmedsattia developmentofanimmunochromatographicstripusingconjugatedgoldnanoparticlesfortherapiddetectionofiklebsiellapneumoniaeicausingneonatalsepsis |