TABLE 7.
PCR-based assays for detection and molecular characterization of intestinal nematodesh
| Species | Amplification method | Detection method(s) | Target(s) | Sample type(s) (no. of samples) | Reference |
|---|---|---|---|---|---|
| O. bifurcum | Nested PCR | Gel electrophoresis | ITS2 | Stool (119) | 486 |
| N. americanus, O. bifurcum | Nested PCRa | Gel electrophoresis | ITS2 | Stool (262) | 13 |
| A. duodenale, N. americanus | Nested PCRb | Gel electrophoresis | ITS2 | Stool (503) | 487 |
| Ancylostoma, N. americanus, O. bifurcum | Multiplex real-time PCR | Hydrolysis probe | ITS2 | Stool (339) | 328 |
| A. lumbricoides | Nested PCR | Gel electrophoresis and sequencing | Cytb | Coprolites (9) | 488 |
| A. lumbricoides | Nested PCR | Gel electrophoresis and sequencing | ITS1, Cytb | Stool (13) | 318 |
| S. stercoralis | Real-time PCR | Hydrolysis probe | SSU rDNA | Stool (370) | 354 |
| A. duodenale, N. americanus, Trichostrongylus | Nested PCR | Gel electrophoresis | ITS1-5.8S-ITS2 | Stool (203) | 489 |
| Trichuris vulpis, T. trichiura | Nested PCR | Gel electrophoresis | SSU rDNA | Human stool (80), dog stool (79) | 311 |
| Ancylostoma, N. americanus, A. lumbricoides, S. stercoralis | Multiplex real-time PCRc,d | Hydrolysis probe | ITS2 (Ancylostoma, N. americanus), ITS1 (A. lumbricoides), SSU rDNA (S. stercoralis) | Stool (1,312) | 319, 321, 333 |
| Ancylostoma, N. americanus, A. lumbricoides, S. stercoralis | Multiplex real-time PCRe | Hydrolysis probe | ITS2 (Ancylostoma, N. americanus), ITS1 (A. lumbricoides), SSU rDNA (S. stercoralis) | Stool (78, 229) | 161, 320 |
| S. stercoralis | Real-time PCR | Hydrolysis probe | 28S | Spiked stool | 353 |
| S. stercoralis (and O. viverrini) | Real-time multiplex PCR | FRET probes-MCA | SSU rDNA | Stool | 356 |
| Ancylostoma, N. americanus, A. lumbricoides, S. stercoralis | Multiplex PCRe | Luminex detection | ITS2 (Ancylostoma, N. americanus), ITS1 (A. lumbricoides), SSU rDNA (S. stercoralis) | Stool (319) | 322 |
| Ancylostoma, N. americanus, A. lumbricoides, S. stercoralis, T. trichiura | Real-time PCRc,d | Hydrolysis probe | ITS2 (Ancylostoma, N. americanus), ITS1 (A. lumbricoides, T. trichiura), SSU rDNA (S. stercoralis) | Stool (525) | 18 |
| T. trichiura, A. lumbricoides | Real-time PCRf | Hydrolysis probe | SSU rDNA (T. trichiura), ITS1 (A. lumbricoides) | Stool | 309 |
| N. americanus | Real-time PCR | SYBR green | ITS2 | Stool (216) | 332 |
| A. duodenale, N. americanus | Nested PCRg | Gel Electrophoresis | ITS2 | Stool (634) | 490 |
| N. americanus, A duodenale, A. ceylanicum, A. caninum, A. braziliense | Real-time PCR | SYBR green-HRM | ITS2 | Stool (634) | 330 |
| S. stercoralis | Real-time PCRd | Hydrolysis probe | SSU rDNA | Stool (160) | 355 |
| Ancylostoma, N. americanus | Real-time PCRc | Hydrolysis probe | ITS2 | Stool (195) | 329 |
a
O. bifurcum primers are described in reference 486.
b
N. americanus PCR primers and probes are described in reference 13.
c
A. duodenale and N. americanus PCR primers and probes are described in reference 328.
d
S. stercoralis PCR primers and probe are described in reference 354.
e
PCR primers and probes are described in reference 319.
f
A. lumbricoides PCR primers are described in reference 319.
g
PCR primers are described in reference 487.
h
Abbreviations: SSU, small subunit; ITS, internal transcribed spacer; MCA, melt curve analysis; HRM, high-resolution melt curve analysis; Cytb, cytochrome b; FRET, fluorescent resonance energy transfer.